PHYSIOLOGYNEWS

summer 2006 | number 63

Meetings UCL Brazil Manchester Munich

Also featuring: Trials and tribulations of self-experimentation Alison Brading’s 10 key papers A century of physiology online Working out aerobic fitness Bursting with currents Granny’s going global

A publication of The Physiological Society 1 MANCHESTER FOCUSED MEETING Frontiers in epithelial transport 6–7 April 2006

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1 Welcome reception, Whitworth Art Gallery 2 David Sheppard (Bristol), left, defends his poster to Seth Alper (Harvard) and Barry Argent (Newcastle) 3 Poster session 4 Welcome reception 5 Lowry Centre 8 entrance 6 Conference dinner at the Lowry Centre 7 The speakers 8 Poster Prize winner Jennifer Evans (Glasgow) 9 Martin Steward (Manchester), 2nd right, 5 shares a joke with Masataka Murakami (Okazaki), 2nd left, and Hiroshi Ishiguro (Nagoya) 10 Jim Melvin (Rochester), left, and Shmuel Muallen (Dallas) 11 Speakers (from left) David Cook (Sydney), Mike Gray (Newcastle) and Karl Kunzelmann (Regensburg) 12 Craig Smith (Manchester), left, and Jeppe Praetorius (Aarhus) at the Lowry Centre gallery 13 Boarding deluxe transport to the conference dinner 14 Conference chair Maynard Case passes round the begging bowl. 7 6

(photos by Austin Elliott, Simon Kellas and Donald Ward) PHYSIOLOGYNEWS Editorial 3 Meetings Physiology at UCL in 2006 John Carroll 4, 15 The Society’s dog. ‘Rudolf Magnus gave Joint International Meeting with the Brazilian Physiological me to Charles Sherrington, who gave me Society José Antunes-Rodrigues 6 to Henry Dale, who gave me to the Images of Manchester inside front cover Physiological Society in October 1942’ Images of Munich 25 IUPS David Eisner, Akimichi Kaneko, Ole Petersen 7 My 10 key papers Alison Brading 9 Published quarterly by the Physiological Society Living history Contributions and Queries My own subject Hugh de Wardener 13 Executive Editor Linda Rimmer Features The Physiological Society Publications Office Targeted cell based delivery of siRNA PR Brink, V Valiunas, S Doronin, P.O. Box 502 Cambridge CB1 0AL I Potapova, RB Robinson, MR Rosen, T Mathias, IS Cohen 16 UK Working out aerobic fitness Ole Kemi, Per Haram, Ulrik Wisløff, Øyvind Ellingsen 18 Tel: +44 (0)1223 400180 Erythropoietin controls ventilation in hypoxia Jorge Soliz, Joseph Fax: +44 (0)1223 246858 Email: [email protected] Vincent, Omolara Ogunshola, Max Gassmann 20 The Society web server: http://www.physoc.org Bursting with currents Wenbo Zhang, Thomas Fisher 22 Developing insights into Purkinje cell maturation Bruce McKay, Magazine Editorial Board Ray Turner 24 Editor Austin Elliott (University of Manchester) Muscle mass at the top: a likely role for fibre hyperplasia in Members humans Giuseppe D’Antona 26 Lesley Anson (Nature) Cardiac channelopathies studied with the dynamic action Patricia de Winter (King’s College London) potential clamp technique Géza Berecki, Antoni van Ginneken 28 Sarah Hall (University of Cardiff) Munir Hussain (University of Liverpool) Reduced mitochondrial efficiency: dysfunction or defence in ageing John Lee (Rotherham General Hospital) muscle? David Marcinek 30 Thelma Lovick (University of Birmingham) Bill Winlow (Prime Medica, Knutsford) Granny’s going global Alister Nicol, Hanno Fischer, Andrew Tate, Keith Kendrick 32 © 2006 The Physiological Society ISSN 1476-7996 Letters to the Editor 34

The Society permits the single copying of Reports individual articles for private study or research. For Goal-directed limb movements Tom Matheson copying or reproduction for any other purpose, written permission must be sought from The ‘Northern’ Cardiovascular Research Group Matthew Physiological Society ([email protected]). Lancaster, Sarah Calaghan 35

Opinions expressed in articles and letters submitted NC3Rs Stakeholder Meeting Tim Watson 36 by, or commissioned from, Members, Affiliates or Moving forward with neuromuscular transmission Colin Ingram, outside bodies are not necessarily those of The Richard Ribchester 37 Physiological Society. The Society’s journals The Physiological Society is registered in England Experimental Physiology Emma Ward 38 as a company limited by guarantee: No 323575. Registered office: PO Box 11319, London WC1X A century (and more) of physiology now online Carol Huxley 40 8WQ Registered Charity: No 211585. Affiliate News Babies and the bench Dola Akanmu 41 Printed by The Lavenham Press Ltd Memorable Members Henry Barcroft Ian Roddie 42 Cover photos Front. The auto-icon of Jeremy Bentham (JB) will be Society News welcoming you to The Physiological Society meeting at UCL in July. In the formation of UCL, JB provided financial Parliamentary and Scientific Committee Ian McGrath 43 support and much of the spirit behind the ethos of what UCL New Chief Executive and President Giovanni Mann 44 would stand for as an institution. Established in 1828 using money raised through selling shares, UCL was originally Biosciences Federation Richard Dyer 45 known as the University of London before being rebranded Unbelievable! 46 as University College London in 1836. The first University established in England after Oxford and Cambridge. It was formed by, what JB described Obituaries as an 'association of liberals' and as such UCL was the first university not to discriminate on the basis of sex or faith (Source: Harte N & North J. The David Greenfield Robert Graham, David Banks, Peter Fentem, world of UCL 1828-2004). Ian Roddie 47 Autar Paintal Tim Biscoe 49 Back. The quad, University College London Book Reviews 51 Noticeboard 52 PHYSIOLOGYNEWS

Action points Guidelines for contributors In this issue Grants These guidelines are intended to assist Welcome to the Summer 2006 edition of For full information on Members’ authors in writing their contributions and to Physiology News, with our usual mix of and Affiliates’ Travel Grants, reduce the subsequent editing process. The features. The scientific articles range Network Interaction Grants, Non- Editorial Board of Physiology News tries to from molecules to man (to borrow a ensure that all articles are written in a Society Symposia Grants, Vacation phrase), including Alison Brading’s 10 key Studentship Scheme, Departmental journalistic style so that they will have an immediate interest value for a wide papers (p. 9) , and a jaw-dropping Seminar Scheme, Centres of readership and will be readable and account from Hugh de Wardener of the Excellence and Junior Fellowships comprehensible to non-experts. In kind of experiments a physiologist could visit: http://www.physoc.org/grants particular, scientific articles should give a carry out on himself back in the days good overview of a field rather than focus before ethics committees (p 13). Membership applications entirely on the authors’ own research. Applications for Full and Affiliate We also report on a couple of notable Membership are received throughout Format of articles announcements. Firstly, The The main message or question posed should be the year and have no deadlines. A introduced in the first paragraph. The background Physiological Society has a new Chief decision is normally made within 7 for the topic should then be established, leading Executive and a new President-Elect days of the Administration Office up to the final conclusion. (p. 44), and secondly the full back­ receiving the application. For full catalogue of The Society’s journals (right Length of articles back to 1878) is now live online (p. 40). details please visit: This will be determined by the subject matter and http://www.physoc.org/join agreed with the Executive Editor. Eagle-eyed readers may spot that this Submission of articles Change of address issue is missing our regular diary feature Authors should submit articles as a Word A Week in the Life. I am sad about this, Members should inform the document attached to an email. Illustrations Administration Office of any changes should be sent as separate attachments (see below) as a diary has appeared in every issue of address, telephone, fax or email and not embedded in the text. since Alan North kicked off the feature in address. PN 55 (Summer 2004). But we had no Illustrations and authors’ photographs volunteers this time, and we finally seem Changes can be emailed to: Authors are encouraged to submit diagrams, to have run out of people that the editorial [email protected] or updated drawings, photographs or other artwork with their articles or to suggest appropriate illustrations. A team have enough incriminating material online at http://www.physoc.org photograph of the author(s) should also on to ‘persuade’ them to become diarists. accompany submissions, if possible. Illustrations However, every threat is another and photographs may be colour or black and white, prints, transparencies or tif/jpeg files with a opportunity (©your head of department), minimum resolution of 300 dpi. Electronic so any would-be diarists out there, get Physiology News colour figures should be saved in CMYK mode. writing. Why not keep a diary of your next conference trip? Or a week of exam Deadlines References Authors are requested to keep the number of season hell? Otherwise I might have to Letters and articles and all other references to a minimum – preferably no more write the next one myself. contributions for inclusion in the than two or three. Please cite all references in the Autumn 2006 issue, No. 64, should style of The Journal of Physiology (see reach the Publications Office Instructions to Authors 2005 at Austin Elliott ([email protected]) by 30 June http://www.physoc.org) 2006. Short news items are encouraged and can usually be included as late copy if space permits.

Suggestions for articles Suggestions for future articles are welcome. Please contact either the Executive Editor or a member of the Editorial Board of Physiology News (see contents page for details).

Physiology News Online Physiology News is now available on The Society’s web site: Oi, ref! http://www.physoc.org. Unbelievable! (p. 46) EDITORIAL PN 3

repeatedly recognise the need for science teaching this process, along with the current and ex­ Scientists needed to be grounded in experimental work. Some of the classroom teachers who presently undertake the pressures reducing the amount of practical science job. It is a mark of many scientists I know that they in secondary (high) schools – overworked attribute their interest in science to two things in teachers, ageing labs and equipment, overstuffed Next there is the question of what experiments their childhood. One is inquisitiveness – posing the test-driven curricula, fears (often misguided) about schoolchildren can actually do. It is not clear to me question ‘why?’ The other is early exposure to the health and safety – have been identified, though how the ‘bank’ of experiments gets renewed, idea that you could sometimes find out why by not solved. except that I see little evidence that working looking at things closely – observation – and/or scientists are involved, which worries me. If we doing experiments – ‘what if?’ This insight into However, I think more work is still needed at the want to come up with new and interesting science experiment as the source of scientific knowledge grass roots from practising scientists – us – and experiments for schoolchildren, fit for the 21st may come from an inspired teacher, from family critically at the institutional level in our century, my starting point would be design teams members, or even from a book, but without it departments, faculties and universities. that should include professional scientists along science will always seem like magic. With it, it can with the experienced classroom teachers and the be recognised correctly as a result of collective One goal should be to de-mystify research and the educationalists. endeavour, thought, and imagination. people who do it, by sending scientists out to schools to make the link between school science So action is needed at all levels. Individual, Everyone, from governments and employers (and the grounding in experimental scientific societal, and institutional. As individuals, there are through to learned societies, agrees we need more method it should provide) and real research. There many ways to get involved. I hope many of us will, science and technology-literate citizens. The are schemes to do this, including the Researchers and perhaps the suggestions above might offer National Curriculum in the UK repeatedly stresses in Residence scheme for PhD students. But much some ways. the need for pupils to be taught to appreciate that more still needs to be done. the scientific facts they learn are based on Learned societies can help, and are helping, both evidence derived from experimentation. And Another problem that I think needs addressing is by lobbying, and by spreading information about experiments have other benefits, too: aiding the distance between professional scientists – the how to get involved. Maggie Leggett, writing in learning – ‘seeing is believing’ – and keeping pupils experts in experimentation and the principles upon Physiology News a year or two back about public interested by making schoolwork interactive and which it is based – and decisions about exactly communication and engagement, suggested that engaging. what practical science is done in school the key role of learned societies was likely to be as classrooms. One might expect that scientific places where people could exchange information But is the message really getting through and professionals would have a close interest in the on what approaches they had tried, what worked, being acted upon? And can the basis of scientific details of how experimental science is first and what didn’t. I am sure there are many knowledge in experimentation really be taught introduced in the early school years, and how it is members out there who have engaged in initiatives while children do less and less hands-on, lab­ taught in the years through to university entry. But to help with science education. But I do not know based experiments? we have been guilty, as I see it, of not getting who or where they are, or what they have done. much involved beyond agreeing ‘there should be Building up web-based registers, talk-boards, and Like many UK academics, I have the impression more of it’. resources to help with this information exchange over the last decade-plus that students starting seems a role tailor-made for learned societies, and university degrees in the biomedical sciences have Part of the reason, I suspect, is that the training of the Physiological Society is increasingly active in done rather little practical work in the later years of science teachers – the key people in helping pupils this regard. secondary school. When I began interviewing comprehend scientific method – is almost prospective undergraduates – final year school completely dominated by educationalists. In case Underlying all this is the role of institutions, and pupils – 10 years ago, one of my standard there is any confusion, an educationalist is defined here I mean, specifically, ‘scientists’ employers’. In questions was ‘what was the most interesting by most dictionaries as ‘a specialist in the theory of particular, universities have to decide whether this experiment you have done at school this last education’. University scientists are scientific is something they care enough about to pay year?’ But I soon stopped asking this, because I educators, often of considerable experience. In the academic staff to take an interest in it, given that only rarely got an answer. Indeed, many could not main, though, they are not educationalists, and do this may take time away from other activities. I remember anything about the few experiments not wish to be. Personally, I feel that professional think it is time for UK universities to grasp this they had done. Strangely, at least to me, many of scientists should be directly involved in the particular nettle. The first step, which many the same students expressed a desire to work in postgraduate training of secondary school science universities and faculties have now taken, is scientific research. teachers. While most people would agree that a appointing specialist personnel tasked with things classroom science teacher needs an like schools liason, community outreach, and So there is a disconnect here. Research as understanding of the theory of education, they also external affairs. But these people are primarily co­ students read about it in the newspapers or in, say, need a good grasp of practical science, its ordinators, and their success will rely critically on New Scientist, seems high-tech and space-age, importance, and how to translate it to the being able to recruit enough willing participants carried out in gleaming labs by modern wizards classroom setting. At present, we assume in the from inside universities. This will depend on pipetting microlitres of mysterious solutions from UK that all the scientific background necessary to institutional leadership, and on recognition of the one tube to another, or operating million pound do this is acquired adequately during a B.Sc. importance of the job to the point where it is seen scanners. Students fail to connect this with the degree course. I remain unconvinced about this, as a core part of at least some scientists’ basics – following a protocol, optimizing a method, especially when the course in question quite responsibilities. careful repetition, observation, formulating and possibly contains no lab-based work beyond the testing a hypothesis – as they themselves may second year. Of course, it is true that teachers We can all agree that ultimately, if enough children have experienced them in school laboratories. The acquire much of their skill and knowledge through are not getting interested in science, there will be a less practical science they do, the more this is true. the process of having to explain things in the gradual decline in the number of high-quality classroom, in much the same way that university undergraduates, and subsequently of scientists. This problem has, to some extent, been physiologists traditionally learnt basic integrative We need action now, particularly from universities, recognised. There have been reports by bodies physiology by demonstrating and explaining it in if this is to be tackled. such as CASE (formerly Save British Science) and lab classes and tutorials. But there is surely a role NESTA, learned societies have voiced their for experienced university teachers in helping concern, and government strategy documents mentor trainee science teachers through some of Austin Elliott

Physiology News | No. 63 | Summer 2006 | www.physoc.org 4 PN UCL MEETING

Physiology at UCL in 2006 Research at UCL Physiology UCL Physiology has a strong reputation in cellular The Physiological Society neuroscience, cell signalling and systems Main Meeting, 4–7 July physiology. Details of the research interests of individual groups can be found on our new web site (www.physiol.ucl.ac.uk). Our research is conducted by 30 academic staff, 11 senior research fellows, 40 post-doctoral scientists and around 30 PhD students. Research activity is flourishing. Members of the Department attract approximately £17 million in research grants from the main UK research councils and charities. An influx of JIF and SRIF Pharmacology provided funding for the Andrew funding has provided nearly £12 million pounds for Huxley Building, which was recently opened by improving the Department’s infrastructure. This has Lord Sainsbury and Andrew Huxley, as well facilitated the bringing together of research groups refurbished labs for five groups. The group has into modern well equipped laboratories. The UCL recently been strengthened by new appointments. tradition of undertaking world class research in Josef Kittler provides a much needed strength in Victorian conditions is a claim we can no longer molecular neuroscience and is interested in the justify. Instead, we have new buildings, refurbished molecular and cellular mechanisms regulating the corridors, and more central facilities to encourage trafficking of GABAA receptors. Troy Margrie and and support interdisciplinary research. Of course, Tom Mrsic-Flogel (arriving in October) will use there is plenty more to do, as some members of the functional in vivo imaging including multiphoton Department will testify, but delegates will notice big microscopy to explore mechanisms of sensory changes since the last Society Meeting at UCL in processing. 2001. Autonomic Neuroscience has been a long Our research falls into a number of defined areas. standing interest in the Department based at The There is considerable and desirable overlap in Royal Free Hospital. Mike Gilbey, Dave Jordan and some areas which facilitates collaboration. We aim Mike Spyer lead groups in this area, focused mostly to use a multi-disciplinary approach that combines on the control of respiration. Alexander Gourine has recently been awarded a Wellcome Trust Senior Fellowship and we are well down the track with plans to move Alex and the Autonomic Group to the It is with great pleasure that the UCL main campus over the summer. Department of Physiology welcomes The Epithelial Physiology Group of Ted Debnam, The Society back to London for its Robert Unwin, Brian King and Scott Wildman are annual Main Meeting. The Department also based at the Royal Free Hospital. Their work is currently busy with final preparations focuses on control of secretion in the kidney and the gut. This group is a joint venture with the to ensure that your visit to London in molecular and bioinformatics techniques with Division of Medicine and will soon be strengthened July will be a veritable feast of science electrophysiology, whole animal recording and with new joint appointments at professor and and social activities. This will be the intracellular imaging using confocal and multi­ lecturer levels. This will provide a strong second of The Physiological Society’s photon microscopy. We have recently expanded in translational angle to the research that will span new style Meetings and we hope that these areas through the recruitment of new staff from molecular genetics of renal transport through to in vivo physiology. London will prove to be every bit as and by investment in new in vivo and in vitro multi­ photon facilities. Dedicated mechanical and exciting as the first, held last year in electronics workshops provide custom-made Sensorimotor Control was the topic of a recent Bristol. equipment that greatly facilitates those particularly Physiological Society Focused Meeting at UCL. difficult experiments. This Meeting was organised by Phil Harrison. Other So far everything is looking good. members of the group include Linda Harrison, Margaret Mayston and John Stephens. The Group Exciting Plenary and Prize Lectures; Cell Signalling Group investigates how cells process the many messages received from the has strong ties with the UCL Institute of Neurology not forgetting the second Physiological external environment. Lipid signalling (Shamshad and with Simon Farmer. Interests include Society Public Lecture to be delivered Cockcroft and Geraint Thomas), and calcium understanding the reflex pathways that control hand by David Attwell, our own Jodrell signalling (Steve Bolsover, Sandip Patel, Jonathan and shoulder movement in man and in patients with specific lesions in the pathway. Professor of Physiology. There will be Gale, John Carroll) are two signalling pathways that we study in a range of physiological systems a number of workshops including including, the immune system, hair cells and oocyte Neuroscience methodologies, Teaching and embryo development. Michael Duchen leads a and Careers in science, as well as 16 diverse multi-disciplinary group that works on the specialist symposia and a total of nearly role of mitochondria in the pathophysiology of cell 400 Abstracts to be presented over the function. three days of the Meeting. The Main Cellular and Molecular Neuroscience Group is Meeting will be preceded by the Young the largest in the Department. Members of this Physiologists’ Symposium, From group have interests in the regulation of ion channel molecules to behaviour on 3–4 July. function (Paolo Pedarzani), synaptic transmission Clockwise from top left: Entrance to Physiology; (Jonathan Ashmore, David Attwell, Angus Silver, All this will provide something of Angus Silver and his new ‘baby’; John Carroll, Frances Edwards), and information processing Tomoko Watanabe, Guilaume Halet and Tasos interest for even the most refined of (Andy Batchelor, Michael Hausser). A JIF bid led by Siskoglou in the new cell physiology labs; Jenga physiological tastes. David Attwell and colleagues in Physiology and Káradüttir in the Andrew Huxley Building.

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Teaching physiology at UCL Physiology at UCL: past present and future The Department teaches physiology to over 1,000 medical and science students. Everyone The Physiology Department at UCL has a in the Department contributes to teaching long and distinguished history that has been either through delivery or organization of entwined with The Society from the outset. courses and modules. There are a core of From its origin in the 19th century, members individuals – Richard Tunwell, Linda Harrison, of the Department (including Burdon- Jonathan Fry, Peter Tatham and Geraint Sanderson, Sharpey, Sharpey-Schafer, Thomas – that run the teaching operation. Starling, Bayliss, Lovatt-Evans, AV Hill, GL Brown, A Huxley, B Katz and many others) The ever-expanding student body has forced have made major contributions to us to take radical steps in how we teach the physiological science. Three of these large numbers of students in the first year. We physiologists, Hill, Huxley and Katz were are indebted to Chris Richards and the awarded Nobel Prizes for their efforts. The Department’s new Teaching Fellow, Pamela role the Department played in fostering the Houston, for transforming our first year discipline of physiology in the UK is illustrated physiology course to include online tutorials by the fact that the first Meeting of The and automatic assessment techniques. We Physiological Society was held in Burdon remain committed to practical classes and Sanderson’s house in College. students continue to learn the experimental nature of physiology in all 3 years. Many also Today the majority of the Department exists opt to undertake a 9 week research project in in the original Institute of Physiology building. their third year. Built in 1909 under Starling’s direction (see Tim Biscoe’s piece below), the Institute The social programme starts with a subsumed the old UCL school playground, Wellcome (sic) Reception on 4 July. perhaps starting a fad that may go some way This will be an opportunity to explore to explain why the UK may excel in the very impressive new home of the physiology and not in events of a more Wellcome Trust. The Society Dinner sporting nature. Developments and mergers on 6 July steps away from the over the years have seen the Department traditional and will take the form of a expand and it now includes the three floors of BBQ in our local green space, the old medical school in the Rockefeller Regent’s Park. Marquees, Pimms, spit­ Building and laboratories at the Royal Free roasted beasts, a string quartet and Hospital. More recently, members of the Department have taken a leading role in perhaps even a fire-eater will make establishing the Ear Institute just east of the this one summer soirée not to be main campus. The Department currently missed (barring, of course, consists of more than 100 members and meteorological interventions – in research interests range from molecular cell which case the marquees will prove physiology right through to in vivo studies snug and popular). In addition to these concerning sensory and autonomic scheduled events the central London neuroscience, regulation of respiration and location of UCL will ensure that there From the top: No worries, we can build it – Duncan motor control (see p. 4 for more on our are plenty of restaurants, pubs and Farquharson and Alan Hogben, Mechanical Workshop; research interests). clubs to cater for every physiologist’s all smiles – Matthew Duckett and Mark Todd in the Teaching Office; new boy, Jo Kittlerr, with members of For the future, physiology will remain a major tastes. his lab – Andrew, Alison, Marzieh, Jo, Chris and Kate; discipline at UCL. However, the university John Carroll Raffaella Bosurgi, Raffaella Tonnini, Paola Pedarzani, structure in which we conduct our research Derek Costello and Ruth Taylor in the new may end up radically different from that which Department of Physiology, University College London, neuroscience labs; Michael Duchen – getting to grips UK with the new multiphoton imaging system. we currently enjoy. Physiologists in the UK Prize Lectures at UCL (p. 15) and beyond will have endured university The original Physiology Laboratory at UCL was found in the North Wing of the main College restructuring and reorganization, with Building, above the Slade School. In the stairwell there is a memorial plaque. Starling departments in Cambridge and Oxford being the most recent. The passion for restructuring appointed as Jodrell Professor in 1899, was dissatisfied with the accommodation and set has now hit UCL and the long term existence about having a large new department, which was completed in 1909 within a year and cost of our traditional departments within a faculty £20,000 to design, build and equip. Starling notes ‘Ironwork outside staircase (Mr T of life sciences is under review. Super­ Elsley)...£135’. There were novel architectural features and, over the front, one can see in gilt faculties of biomedicine and health and letters Institute of Physiology in art nouveau style. This was a tribute to the German fashion disease-driven institutes are the order of the admired by Starling, and ‘much disliked by the then Provost, who thought he scented day. Perhaps it is time to accept the secession’ (Lovatt Evans,1964). inevitable cycle of fashion and reinstate the Tim Biscoe ‘Institute of Physiology’, perhaps with a Honorary Member splash of ‘integration’, ‘translation’ and a good Lovatt Evans C (1964). Reminiscences of Bayliss and Starling. Cambridge, The University Press. dose of medicine.

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Joint International Meeting with the Brazilian Physiological Society

For the last 2 years the Organizing and Scientific Committees have been directing efforts to ensure that the Meeting will attract scientists – both faculty and students – dedicated to all areas of physiology. The programme has been carefully planned to allow the attendees to find up-to-date information as well as providing time to discuss and debate science with their peers. Congresses offer an excellent opportunity to meet one’s professional friends, be introduced to others, and share different opinions and cultural perspectives.

The Convention Centre of Ribeirão Preto, venue of the Congress, provides the latest high technology facilities. It is located in the heart of the town, close to many hotels and commercial areas. The airport can be reached within 15 minutes from the city centre by car. Ribeirão Preto, 319 km from São Paulo, will host the 41st Congress of the The Brazilian Physiological Society, as Brazilian Physiological Society from the local host, is looking forward to 27 to 30 August. It will be held jointly welcoming you to Ribeirão Preto in with The Physiological Society. Since August and to chatting over a cup of both partners seek the advancement of good Brazilian coffee! physiology as an integrative science aiming at the understanding of the A history of the Brazilian mechanisms that explain how the body Physiological Society works, a most exciting and productive The Brazilian Physiological Society encounter can be anticipated. (SBFis) was established in 1957 by the initiative of some of our country’s most eminent physiologists in an effort to promote physiology, encourage interchange of ideas and to improve the opportunity of young researchers to meet and discuss their projects and ideas with eminent physiologists.

Since then, Congresses of the SBFis have been held every year and the President, Secretary and Treasurer elected every 3 years. From the top (above): the Metropolitan Cathedral of In the past 20 years the SBFis has São Sebastião in Ribeirão Preto, located at Praça das established numerous scientific Bandeiras; Francisco Schmidt Coffee Museum in the commissions in various specialised botanical gardens of the Historical Museum; Buzios- areas of physiology, such as Rio; map showing the location of Ribeirão Preto; current SBFis President Walter Zin. undergraduate and graduate teaching and scientific committees. SBFis also Left: Petropolis, near Rio (top) and Bahia-Salvador.

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International Union of Physiological Sciences

I hope that Members of The Physiological Society will be particularly interested in the future of the IUPS, not least for the reason that The Society will be hosting its 2013 Congress. Members are encouraged to send their views on IUPS to the Long­ range Planning Committee.

The article mentions the IUPS Africa Initiative. This was mentioned in a previous issue of Physiology News (59, 10) organised a number of symposia and Top: Christ the Redeemer, Rio de Janeiro; and several Members of The Physiological regional meetings, as well as advised Above: Sunset by the riverside in Belem do Para in Society have kindly written to offer help. the Scientific Program Committee of the delta of the Amazon River. We are currently trying to organise plans the Federation of Societies for The main goals of the SBFis with both African and IUPS colleagues. Experimental Biology. • to stimulate research leading to the Finally, links with the IUPS can only be progress, diffusion and knowledge of In 1956, 1961 and 2003 the SBFis physiological sciences further improved by the recent organised joint meetings with the Latin announcement that its Secretary General American Association of Physiological • to stimulate the improvement of (Ole Petersen) will become the next Sciences, when it was given the methods to teach physiological sciences President of our Society (see p. 44). opportunity to promote the interchange according to modern technologies amongst eminent Latin American and David Eisner • to promote remarkable training of International Secretary other scientists from abroad, young physiologists in order to strengthen the graduate and undergraduate students, physiological sciences in our country The following update on IUPS activities interchange of experiences, as well as and abroad is adapted, with permission, from the the establishment of scientific first of a newly established series of cooperations among Brazilian • to encourage the young scientist to be IUPS Editorials in Physiology (21, 2). physiologists. The last joint meeting of involved with the understanding of the ALACF with the SBFis – systemic physiology, combined with The most important recent IUPS event Challenges for the advance of new technological developments was the 35th International Congress of physiology in the third millennium – (molecular, genetics, immunology, Physiological Sciences held in San was held in Ribeirão Preto, São Paulo, imaging, etc.) Diego, California, from 31 March to 5 Brazil, and was attended by 1244 April, 2005. This Congress had many • to promote the integration between international scientists. new features, including a very success­ basic and applied research ful series of tracks covering some of • to support the participation of young the most important current Current Officers of the Brazilian scientists and students in the meetings physiological themes. As part of Physiological Society Experimental Biology, there were • to promote cooperative work between Walter Araújo Zin (President) ample opportunities to benefit from, research groups. Francisco Tadeu Rantin (President-elect) and participate in, events organized by Patrícia Rieken Macêdo Rocco (Secretary) José Antunes-Rodrigues friendly neighbouring subjects and for Denise de Carvalho (Treasurer) Brazilian Physiological Society

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members of these other societies to see IUPS 2009 International to Denis Noble via the IUPS Secretariat what physiologists had to offer. With Scientific Programme in Paris (e-mail: [email protected]). more than 10,000 registered Committee participants, this gave unprecedented Chair • Council also elected a committee to exposure to physiological research at a Yoshihisa Kurachi (Japan) come forward with specific plans for an time when this subject is undergoing a Africa Initiative. Tony McKnight is major revival. Vice Chair chair, with Ann Sefton and David Ole H Petersen (UK) Oyebola as members. The Committee is The 35th IUPS Congress in San Diego Ex officio members expected to develop a strategy to be provided a much needed replenishment Akimichi Kaneko (Japan) presented to a funding agency to of IUPS resources. We therefore have Pierre Magistretti (Switzerland) culminate in a major workshop in good reasons to be very grateful to the Africa in 4 years. Council has International) members appointed allocated seed money to get this American Physiological Society (APS), by the IUPS not only for organizing a superb scien­ Yung Earm (South Korea) potentially important initiative under tific event, but also for helping the Malcolm Gordon (USA) way. IUPS rebuild our finances so that we John Hall (USA) • after 4 years of working with the may again plan major initiatives. As Cecilia Hidalgo (Chile) Commission Structure established by described in the 2004 and 2005 IUPS Hans Hoppeler (Switzerland) the 2001 Council elected in Newsletters (http://www.iups.org), the Peter Hunter (New Zealand) Christchurch, New Zealand, it was scientific programme for the San Diego Caroline McMillen (Australia) Ole Petersen (UK) decided to make some adjustments. Congress was generated by the IUPS Quentin Pittman (Canada) Based on the experience of planning for International Scientific Program Irene Schulz (Germany) the San Diego Congress, it was felt Committee (ISPC) in cooperation with Ann Sefton (Australia) desirable to have a new Commission on the APS under the chairmanship of Curt Sigmund (USA) Molecular and Cellular Biology. The Walter Boron. Walter did a fantastic job two Commissions that had previously for which IUPS is extremely grateful. Local Programme Committee Members dealt with various aspects of the Ken-ichi Honnma, Yoshihiro Kubo, Harunori neurosciences were merged into one The San Diego Congress also repre­ Ohmori, Hideyuki Okano, Yasuo Sakuma consolidated Neurobiology sented the crowning achievement of Makoto Suematsu Commission. The new Commission IUPS President Allen Cowley, Jr. Allen Chairs are listed on our web site. was an outstanding President, responsi­ Associate Members Yukiko Gotoh, Tadashi Isa, Fumihiko Kajiya, ble for a major restructuring and Yoshikatsu Kanai, Kazuyuki Kanosue, Itsuo Although it may seem early, planning streamlining of the Union’s Commis­ Kodama, Masato Konishi, Osamu Matsuo, for the 2009 IUPS International sion Structure, which did much to facil­ Katsuhiko Mikoshiba, Hiroshi Nose, Masahiro Congress of Physiological Sciences is itate the scientific programming process Sokabe, Miyako Takaki, Jun Tanji, Gozoh already under way. The 36th for the San Diego Congress. Tsujimoto, Tadashi Tsumoto, Toshihiko Yada, International Congress of Physiological During the San Diego Congress, the Megumi Yoshimura Sciences will be held in Kyoto, Japan, IUPS Council (for composition see the from 27 July to 1 August 2009. IUPS web site mentioned above), The IUPS ISPC has already been chaired by Akimichi Kaneko, who at activities, as well as their resource established (see box, left) and the first the General Assembly in San Diego implications, to the IUPS Council when meeting took place in Osaka, Japan, on was elected to succeed Allen Cowley as it meets again in 2007. The LRPC is 20–21 January. At this meeting the IUPS President, met several times and a now seeking opinions from member fundamental policy for the generation number of important decisions were societies and/or individuals on the of the scientific programme was made. The following three are perhaps future of the physiological sciences, the established and a start made on the of particular significance: organization of IUPS, IUPS meetings, selection of Plenary Lecturers. Some finance, relations with the International details about the Congress are already • a Long-Range Planning Committee Council for Science (ICSU), the United available at the IUPS web site. (LRPC) was established, under the Nations Educational, Scientific and chairmanship of Denis Noble, who Cultural Organization (UNESCO), The IUPS Executive Committee also served as IUPS Secretary General from International Brain Research met in January and we hope to give 1993-2001, when Ole Petersen took Organization (IBRO), and other information about the decisions made at over. Denis has since completed the international organizations, as well as these two meetings in due course. composition of this key Committee. We on public relations. are grateful to Denis, Allen Cowley, Akimichi Kaneko Cecilia Hidalgo, and Yasunobu Okada We are very interested in receiving President opinions from a wide range of the sci­ for taking on this task. The Committee Ole H Petersen will present its report, making entific community. Anyone wanting to Secretary General recommendations on future IUPS comment on these issues should write International Union of Physiological Sciences

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computers and no calculators – a My 10 key papers mammoth task! The memorable papers that Alison Brading (right) still 2 Bülbring E (1954). Membrane potentials of smooth muscle vividly remembers first fibres of the taenia coli of the reading guinea-pig. J Physiol 125, 302-315 This paper was of great importance to For my top 10 papers I have decided to my future. During my PhD (a include those that I still remember first microelectrode study of the body wall musculature of the pig round worm, reading, and which for me were Ascaris lumbricoides) under the memorable, so much so that for most of supervision of Peter Caldwell (who them, I can actually recall where I was used to join the other physiologists who when I read them. Most of the early ones worked on squid axons in the came from The Journal of Physiology, months I had to drive 2 miles to find a Plymouth Marine Biological Centre at because I was a Member of The Society lavatory I could use!). Under the Plymouth in the summers), I was early on (and later on the Editorial Board) chairmanship of John Harris, the course recording spontaneous electrical and had time then to read anything in it was a good traditional romp through activity of a sort I had never seen in the that interested me. Only a few of these the animal kingdom, but with an literature, until I came across papers by papers bears directly on my own unusually large component of Edith Bülbring’s group. I saw to my research, but I am sure they influenced physiology. We were introduced to the amazement recordings of potentials me indirectly, and all gave me Hodgkin and Huxley model of the very similar to mine (see Fig. 1). considerable pleasure. I realise now how action potential in the squid giant axon. lucky I have been in my life as a scientist, I enjoyed a superficial acquaintance As so little was then written about because I eventually met virtually all of with the papers then, but later in my nematode muscle, I followed the the authors, with the exception of David capacity as tutor for physiology at Lady smooth muscle literature, which in this Marr. Looking at the selection I can see Margaret Hall, Oxford, I read the country nearly all emanated in Oxford that, although I started academic life as a papers more carefully, and began to from Bülbring’s laboratory. The paper really appreciate how beautifully zoologist, and came to think of myself as I have included is one I read later, and simple and wonderfully predictive the only with hindsight did I realize what primarily a physiologist, the fact that I Hodgkin Huxley model was. Hence an amazing achievement this work was. have worked entirely in a pharmacology my inclusion of this outstanding paper Microelectrodes had not long been department has clearly had a significant in my top 10 papers. It still pleases me, invented, and to record simultaneously impact! I will refer to the papers in their and I love pointing out to my students membrane potential and contractile approximate date order. how each reproduction of the action activity from strips of smooth muscle potential required iterative solving of composed of cells only a few microns 1 Hodgkin AL & Huxley AF (1952). numerous different equations for every wide and moving spontaneously, was a A quantitative description of membrane current and its point, at a time when there were no stupendous achievement, not even application to conduction and excitation in nerve. J Physiol 117, 500-544 A This paper I came across as an undergraduate. I was fortunate to do my first two degrees in what was then (I started in 1959), probably the best zoology department in the country. Going to Bristol University was largely serendipitous, because having missed 2 years after leaving school (through B contracting polio) I was loath to waste another year getting my A- level work up to university entrance examination standard, which eliminated London, Oxford and Cambridge from consideration. Reading turned me down, but Bristol didn’t seem to mind me being in a wheelchair, even though Figure 1. Comparison between action potentials in a nematode and a mammalian smooth muscle. the layout of the department was totally A, Ascaris lumbricoides body wall muscle (Brading thesis, 1965). unsuited to the disabled (for the first 6 B, Guinea-pig taenia caeci (Bülbring, 1955).

Physiology News | No. 63 | Summer 2006 | www.physoc.org 10 PN MY 10 KEY PAPERS matched that often in later work on in which the cells were electrically that he was able to convince them of smooth muscles. I became deeply interconnected in three dimensions (see their basic error, and to do it in a interested in the smooth muscle work, Fig. 2). Tomita became one of my manner that kept a friendly relationship and gratefully took up the opportunity heroes. An enormously intelligent between the groups. to join Bülbring’s group as a postdoc, scientist with a background in physical hence sealing my future, and gaining in sciences, and a superb experimentalist, 4 Marr D (1969). A theory of her a beloved mentor. he was nevertheless a modest man, and cerebellar cortex. J Physiol 202, hated confrontation. It was difficult to 437-470 3 Tomita T (1966). Electrical persuade him to interpret his results in The next paper on my list has nothing responses of smooth muscle to his papers, because he believed that if to do with smooth muscle, but hit my external stimulation in hypertonic he just gave his results everyone else attention when reading the then current solution. J Physiol 183, 450-468 would be intelligent enough to see the issue of The Journal. I remember When I first joined Bülbring’s implications. At that time another coming across it in the departmental laboratory Tadao Tomita was working active group of smooth muscle library and staying there to read it with her. This ground-breaking paper, researchers were attempting through, which took me several hours! published soon after I arrived, had an erroneously to measure membrane This was a real ‘eye opener’ and enormous influence on research into the properties in smooth muscle tissues by extremely exciting. It was the result of properties of smooth muscle. Using a injecting current into single cells Marr’s work during the tenure of a long strip dissected carefully from the through a microelectrode, which research studentship at Trinity College guinea-pig vas deferens, and a partition Tomita’s work had clearly shown to Cambridge. Based largely on the stimulating method to apply a uniform give grossly inaccurate results. detailed anatomical findings of Eccles, change in potential to the cells at the Marr put forward a remarkably partition, Tomita measured the spread Because this group were influential, we complete theory of how the cerebellum of the membrane potential change with tried to persuade Tomita to discuss their could learn to perform motor skills, and distance from the partition using work with them, but he would not. made some predictions. The most microelectrodes, and showed the then However, at an International meeting in important prediction was that ‘the extremely surprising result that cable Munich their group were seated at a synapses from parallel fibres to theory predicted the results, enabling table close to ours at the main dinner. Purkinje cells are facilitated by the the time and space constants of the On the table also were quite large shots conjunction of presynaptic and tissue to be derived. For those of schnapps for each delegate, and the climbing fibre (or post synaptic) interested, a detailed theoretical model beer flowed freely in litre steins activity’ in other words behaved as is described in Tomita, 1966 and shows throughout the meal. Playing on the suggested earlier by Hebb (1949). In how the membrane resistance and common difficulty in metabolizing spite of the fact that evidence now capacitance could be measured. The alcohol that many Japanese have, we suggests that the conjunction of activity implication of the paper was that the waited until Tomita’s inhibitions were actually depresses rather than facilitates smooth muscle cells in the vas deferens sufficiently blunted, and pointed him in response to parallel fibre input, this were composed into functional bundles their direction. I am delighted to say paper for the first time gave the feeling

Figure 2. The effect of hyperpolarization on the action potential of guinea-pig vas deferens. Conditioning stimuli were applied at B, the membrane potential was measured with a microelectrode (NE) near the partition, and action potentials were evoked by the electrodes at A. From Tomita, 1966.

Physiology News | No. 63 | Summer 2006 | www.physoc.org MY 10 KEY PAPERS PN 11 that it might be possible to understand two state model for directly coupled microelectrode voltage clamp technique neural processing. The thinking of this receptors, with a binding event and an to measure currents through nicotinic remarkable scientist and mathematician isomerization. Interestingly, however, receptors activated by various agonists, continues to have a major influence in as pointed out by Andersen and Stevens at a range of different holding understanding of CNS processing in (1973) in The Journal later the same potentials, and then to examine the spite of his tragically short career (he year this model predicts the channel effects of antagonists. I very much died of acute leukaemia in Cambridge, open duration would vary about a liked the approach taken in the paper, Massachusetts at the age of 35). mean, rather than being a ‘shot’ event, and the use of various techniques such and using this assumption these authors as voltage jump etc. to build up a 5 Katz B & Miledi R (1972). The calculated a mean channel open time convincing picture of how the statistical nature of the some five to six times longer. After his antagonists were working. In particular acetylcholine potential and its retirement, Katz was an Editor of The it showed clearly the difference molecular components. J Physiol 224, 665-699 Journal during my tenure as a between the drugs that blocked through Distributing Editor, and was a godsend. classical competitive antagonism and I had long been a fan of Katz and He dealt with his mail in order each those that were non-competitive. It Miledi and read most of the earlier day, and when he came across a paper provided excellent evidence that these papers on the neuromuscular junction. to review, did it immediately and sent it non-competitive drugs were acting as This one published in The Journal took back by return of post! channel blockers. As far as I am aware a huge step forward. For me it was this is the first and only time anyone particularly exciting, because it threw 6 Ascher P, Large WA & Rang HP has clearly demonstrated how a non­ light on the behaviour of the nicotinic (1979). Studies on the mechanism competitive receptor antagonist is receptors, and I had become interested of action of acetylcholine working. Interestingly tubocurare, in receptors (through coffee time antagonists on rat which at the neuromuscular junction discussions with Humphrey Rang and parasympathetic ganglion cells. J Physiol 295, 139-170 does act as a competitive antagonist, at Jim Ritter). This unusually long (for ganglia acts predominantly as a channel them) and detailed paper arose from the This is another paper I came across in blocker, as does hexamethonium, observation that bath applied The Journal. Rang had left the which has little effect at the acetylcholine generated not just department by then for a transient neuromuscular junction. This depolarization, but an increase in the period at St George’s, but I followed highlights the differences between noise of the recording. In a lecture he his research, and this paper struck me nicotinic receptors at the two sites, and gave to The Physiological Society after as a good one. Taking advantage of the sheds further light on the channel his retirement, Katz told how they had simple morphology of ganglion cells, properties. to change their tissue bath in order to the authors were able to use a two administer exogenous acetylcholine (previously they had just studied nerve evoked responses). With the new bath, they initially thought that that the noise that always occurred when they opened the tap to allow the drug access, was spurious, and they spent many hours trying to find the external source, before eventually realizing it was in fact a genuine phenomenon, and would be expected to give information about the underlying molecular events. They chose to analyse the noise (initially by projecting traces onto the screen of the UCL physiology lecture theatre and measuring things with a ruler!) with the assumption that it was caused by random occurrence of ‘shot’ events caused by opening of the channels for a particular duration, and estimated this from their analysis as being of 1 ms. The two state model they proposed, namely:

with two forward and two backward Figure 3. Patch clamp recording of single nicotinic acetylcholine channel activated by four different agonist drugs. rate constants is now a commonly used Adapted from Colquhoun & Sakmann, 1985.

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The last four papers were not published in respond to bath applied agonists, the affinity (the ability of the agonist to The Journal, but came to my attention in lack of specialized endplates and the bind to the receptor) as separate other ways. Continuing with the theme of apparent rarity of nerves in EM entities. Probably not. I have been channels and receptors, for a period indirect sections of many smooth muscles), had fortunate in knowing this eccentric methods such as noise analysis and voltage to be revised, since it was now possible brilliant pharmacologist, and am jumps became the methods of choice for to record the invading action potentials grateful for his patience for the many studying channel properties, until, that is, at the same time as any subsequent times when I ring him up asking him to the coming of the patch clamp. currents resulting from transmitter explain some point or other abut release on the smooth muscle. What receptors. This is a long paper, but full 7 Neher E & Sakmann B (1976). these authors clearly showed was that of insight and wisdom. Single-channel currents recorded in the varicose nerve terminals release from membrane of denervated of a quantum of transmitter from a 10 Gabella G (1995). The frog muscle fibres. Nature 260, structural relations between 799-802 release site was a rare event (as low as only 1 release for 100 action potentials nerve fibres and muscle cells in Few people interested in channels can the urinary bladder of the rat. J when the frequency of stimulation was have missed this paper and will Neurocytol 24, 159-187 low) and caused a discrete transient remember the great excitement the This last paper is relevant to the response of the innervated cell. The possibility of recording currents research area in which I have spent intermittency was not caused by failure through individual channels generated. most of my time in recent years, that of of the action potential to invade the I am going to cheat here and include the control of the urinary bladder. I terminals. It was now possible to Hamill et al. (1981 ), the paper in met Giorgio through Edith Bülbring, compare quantal release from which the improved giga seal method and he has also become one of my autonomic nerves, with that at the was described. This method allowed heroes. He is clearly a brilliant NMJ, and also with individual synapses visualization of the opening and closing microscopist, but his work stands out in the CNS, (being studied at that time of individual channels in a wholly because it is always designed to in Oxford by Julian Jack’s group). The satisfying way, giving definitive proof illuminate some key physiological or ability to study release from single of the underlying theory of the agonist functional point. I have spent many release sites has of course allowed huge generated noise. I am constantly happy hours pouring over his advances in understanding of how amazed and delighted that we electronmicrographs gaining insights synaptic release is modulated, and links physiologists can follow the behaviour into the tissues and their functions. In up with the Marr paper and the of single molecules in real time, with this paper, to study in detail the expanding fields of study into long­ comparatively simple and cheap relationship between the nerves and term depression and facilitation in the apparatus whilst the biochemists with smooth muscle he cut serial sections for CNS. their enormously expensive technology electron microscopy to reconstruct the are still looking at populations! (see varicosities. This is a technical tour de 9 Colquhoun D (1998). Binding, Fig. 3) gating, affinity and efficacy: the force, and the time, effort and patience interpretation of structure-activity to do this work is something few 8 Brock JA & Cunnane TC (1987). relationships and of the effects of possess. Single sections do not allow Relationship between the nerve mutating receptors. Br J the real density of the innervation to be action potential and transmitter Pharmacol 125, 924-947 appreciated, but this detailed analysis release from sympathetic Maybe I am now a pharmacologist! postganglionic nerve terminals. shows that each smooth muscle cell Nature 326, 605-607 This paper impressed me enormously, receives at least one close contact with and addresses a fundamentally This paper describes another a naked varicosity. The images are pharmacological problem, addressing breakthrough arising from application superb, and the results consistent with the issue of whether or not it is possible of a simple experimental approach, that the growing belief that in many cases to measure efficacy (the ability of an of recording activity from underlying the autonomic nervous system does agonist to activate a receptor) and nerves and smooth muscle cells in the indeed make close contact with its effectors. vas deferens using an extracellular Other references glass electrode – a sort of loose patch. Anderson CR & Stevens CF (1973). Voltage clamp analysis of This approach brought about a huge acetylcholine produced end-plate current fluctuations at frog The fact that my top ten papers end in increase in our understanding of neuromuscular junction. J Physiol 235, 655-691. 1998, does not mean that I have not read autonomic neuromuscular transmission, Colquhoun D, Sakmann B. (1985) Fast events in single-channel more recent memorable papers, but that currents activated by acetylcholine and its analogues at the frog none of them have been memorable particularly from sympathetic muscle end-plate. J Physiol 369, 501-557. adrenergic nerves, and began to enough for me to remember where I was Hamill OP, Marty A, Neher E, Sakmann B & Sigworth, FJ (1981). when I first read them, or maybe this is just revolutionize thinking. The general Improved patch-clamp techniques for high-resolution current view that autonomic nerves released recording from cells and cell-free membrane patches. Pflügers a reflection of lack of available memory transmitters that diffused towards the Archives 391, 85-100. space in my ageing brain! underlying smooth muscles, and bathed Hebb D (1949). The organization of behaviour. Wiley, New York. them in transmitters (a view based on Alison F Brading Tomita T (1966). Membrane capacity and resistance of the ease with which smooth muscle mammalian smooth muscle. J Theor Biol 12, 216-227. Department of Pharmacology, University of Oxford, UK

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My own subject say ‘I can’t see anything’ and then ‘I feel sick’ and I passed out. As I did so Hugh de Wardener recalls I am told that I vomited onto the back the trials and tribulations of and head of the ward sister who grimly self-experimentation in the stuck to her task. The results showed 1950s that during the low arterial phase of the faint there was evidence of renal vasodilatation with little change in In the 1950s I worked in Professor renal blood flow. Subsequently, we Peter Sharpey Schafer’s Department of kept the subjects horizontal and pooled Medicine at St Thomas’ Hospital, and the blood in the legs with thigh cuffs, was one of the experimental subjects in which proved more experimentally Hugh de Wardener (right) pictured with Adetokunbo three experiments on renal function. Lucas at the Imperial College Faculty of Medicine tractable. annual award of fellowships ceremony in 2004.

In the first of these, we were measuring In the second experiment we studied the effect of a vasovagal faint on renal the effect of solute secretion on the the nature of the osmolality of the fluid blood flow (de Wardener & McSwiney, production of a hypotonic urine in man that flows into it. 1951) (Fig. 1). Fainting was induced (de Wardener & del Greco, 1955). The by ‘pooling’ blood in the legs followed point in contention was whether the I decided to explore the effect of an by a venesection from an arm vein. I tubular fluid delivered to the distal osmotic diuresis in the absence of was the first subject and the only one in concentration site was isosmolar or endogenous vasopressin. As a whom pooling of blood in the legs was hypoosmolar. It was well kown that the preliminary experiment I decided to see accomplished by sitting on a saddle on induction of an osmotic diuresis the effect on my urine osmolality of a tilting table set at 45 deg. with my resulted in a hypertonic urine which inducing an osmotic diuresis, after legs unsupported. A steady flow of asymptotonically approached having inhibited my hypophysis from urine was ensured by intravenous isosmocity. But to induce an acute secreting ADH with one large oral mannitol. Urine was collected via a osmotic diuresis it is necessary to give intake of alcohol. Once again I had a urinary catheter into small conical an intravenous infusion of hypertonic urinary catheter in place and two flasks firmly kept in place manually by fluid. This stimulates anti diuretic intravenous lines. As we were short­ the ward sister who had volunteered to hormone secretion thus raising the level staffed at the time, I was also writing help. As the venesection proceeded I of plasma ADH, which influences the up the notes of the experiment as it suddenly lost my sight. I heard myself distal concentration site thus obscuring went along. After several control periods, I drank 100 ml of 100% ethyl alcohol in about 1 minute. I came across the notes of this experiment a few years ago and, to my surprise, found that after drinking the alcohol the notes are written with a red biro. After some minutes there is a note ‘I feel like singing’ and a few minutes later ‘Am singing’. The mannitol infusion was then given and the osmotic diuresis induced. Within a short time the combination of alcohol and mannitol gave me a terrible headache. But the results were encouraging and we eventually explored the problem in patients with diabetes insipidus in whom it was unnecessary to give alcohol.

The results were consistent with Shannon’s hypothesis that the fluid entering the concentration site in the kidney is always hypotonic. One of the best demonstrations in favour of this hypothesis is illustrated in Fig. 2. The hypotonicity of the urine of a patient Figure 1. The effect of vasovagal faint on renal blood flow, showing the fall in renal resistance which occurred with diabetes insipidus was gradually with the onset of hypotension (from de Wardener & McSwiney, 1951). reduced by a continuous infusion of

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diminution in the number of aquaporins mobilised to the surface of the collecting tubule cells (Knepper, 1998).

Drinking that much water was most unpleasant. The tediousness and repetitiveness of overcoming one’s increasing distaste for water was exhausting. It is very difficult to drink large amounts of water, day after day, when one is not thirsty. The large volume can only be managed by spreading the intake throughout the 24 hrs, so alarm clocks are needed at night. The most striking subjective feature was a feeling of coldness associated with pallor which varied in intensity. Emotional lability was pronounced, with bouts of hilarity alternating with moroseness. Appetite, however, was excellent throughout, Figure 2. Effect of intravenous pitressin and concentrated mannitol on the urine osmolarity, solute output and urine flow. The urine osmolarity changed from hyper- to hypotonic upon the administration of mannitol (from De though the food was tasteless unless Wardener & Del Greco, 1955). one added large amounts of salt, and the very thought of adding salt caused pitressin until it became mildly Not surprisingly, I had difficulty profuse salivation. This was associated hypertonic. At this point, as the finding volunteers until Andrew with a sharp fall in salt taste threshold pitressin infusion continued, an osmotic Herxheimer’s curiousity about the which gradually returned to critical diuresis was induced by intravenous effect of overhydration on saliva levels by the end of the period of high mannitol, and the hypertonic urine duly induced him to join me. We decided to water intake. became hypotonic. drink 10 l of water per day for 10 days. The effect on the kidneys’ ability to After a week, to relieve the tedium, The third experiment in which I was a concentrate the urine was clear cut (Fig. Andrew and I went over Lambeth subject was the most arduous (de 3). After 10 days the ability to Bridge to the Tate Gallery. The floor at Wardener & Herxheimer, 1957a; de concentrate had fallen from about 1,000 the back of the Morris Minor was filled Wardener & Herxheimer, 1957b). In m.osmol/kg to approximately 500 with numerous large glass containers, the 1950s, when it was not possible to m.osmol/kg. Many years later this some empty for urine collection and measure vasopressin, it was often phenomenon was shown to be due to a others containing drinking water. difficult to distinguish diabetes insipidus from compulsive water drinking (psychogenic polydipsia). A period of dehydration was supposed to separate the two since the concentration of the urine of the compulsive water drinker, who has an intact hypophysis, should then resemble that of a normal person, i.e. it should be substantially higher than that from a patient with diabetes insipidus. I was impressed, however, by the relatively poor ability of patients with compulsive water drinking to concentrate their urine after a period of dehydration, and I wondered if this might be due to a functional adaptation of the kidney to the excess water intake. In order to determine whether this was true, it was necessary to measure the kidney’s ability to concentrate the urine before Figure 3. Urine concentrations following the intravenous administration of pitressin, and after a 26 hr period of and after a relatively prolonged period fluid deprivation, before, and at the end of, drinking about 10 l of water a day for 11 days. The plasma osmolarity of overhydration. varied between the limits indicated by the two lines (from de Wardener & Herxheimer (1957a)

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Prize Lectures The following Prize and Plenary Lectures have been scheduled to take place at the Main Meeting of The Physiological Society at UCL from 4–7 July.

The Physiological Society Public Lecture – David Attwell (UCL) The energetic brain Tuesday, 4 July at 1830

Biller Prize Lecture – Matthew Bailey (University of Edinburgh) Mechanisms of hypertension in a mouse model of apparent mineralocorticoid excess Wednesday, 5 July at 1200

Sharpey-Schafer Lecture – Neville H McClenaghan (University of Ulster) Physiological regulation of the pancreatic beta-cell: functional insights for understanding and therapy of diabetes Wednesday, 5 July at 1400

Hodgkin-Huxley-Katz Prize Lecture – Thomas J Jentsch (Universität Hamburg) Hugh de Wardener (1960) (top left); Peter Sharpey-Shafer (top right) and Andrew Herxheimer (above) Roles of vesicular CLC chloride transport insights from disease, mouse models and biophysics Stupidly we had only labelled the sides to concentrate the urine, we didn’t Wednesday, 5 July at 1815 of the containers, and not the tops, so drink anything for 24 hr. This last that they were difficult to identify. One period seemed endless – we were Joan Mott Prize Lecture – Susan at a time we would come out of the disgruntled, unreasonable and Wray (University of Liverpool) museum, sit in the back of the car, cantankerous. The results, however, Insights into the uterus drape a rug over our knees and empty cheered us up no end. Thursday, 6 July at 1400 Hugh de Wardener One at a time Andrew and I Wellcome Prize Lecture – Helen Emeritus Professor of Medicine, Charing Cross would sit in the back of the Kennedy (University of Bristol) Hospital, London and Fellow of the Faculty of Detecting sounds: mechano-electrical car, empty our bladders, Medicine, Imperial College, London, UK transduction in the mammalian drink some water and cochlea References re-enter the museum De Wardener HE & McSwiney RR (1951). Renal haemodynamics in Friday, 7 July at 1400 vaso-vagal fainting due to haemorrhage. Clin Sci 10, 209-217. our bladders into a container, drink De Wardener HE & del Greco F (1955). The influence of solute secretion rate on the production of a hypotonic urine in man. Clin some water and re-enter the museum. Sci 14, 715-723. It wasn’t long, however, before we De Wardener HE & Herxheimer A (1957a). The effect of a high began to have difficulty distinguishing water intake on the kidney’s ability to concentrate the urine in man. the containers that held the drinking J Physiol 139, 42-52. water from those that contained clear De Wardener HE & Herxheimer A (1957b). The effect of a high hypotonic urine, and we had to go back water intake on salt consumption, taste thresholds and salivary secretion in man. J Physiol 139, 53-63. to the lab. Knepper MA (1998). Long-term regulation of urinary concentrating At the end, to test the kidneys’ ability capacity. Am J Physiol 275, F332-F333.

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Targeted cell-based delivery of siRNA Small interfering RNA (siRNA) is an ever more popular tool for silencing gene expression. But do experimenters always have to deliver siRNA directly into all the cells? Or can siRNAs pass from one cell to another via gap junctions? Ira Cohen and friends suggest that it all depends what connexins are around The only direct form of intercellular (Goldberg et al. 2004;Tsien & pair in which junctional conductance communication in an ensemble of Wiengart, 1976). Although, Kolodny and fluorescence intensity of the tagged animal cells is provided by gap junction (1971) claimed that mRNA was oligonucleotide are simultaneously channels. All other forms of cell-to-cell permeable to gap junction channels his monitored over time (Valiunas et al. communication must utilize the observations were compromised by a 2005). Figure 1A illustrates the transfer extracellular space. In vertebrates there lack of controls for the actions of of a morpholino oligonucleotide 12 are two unrelated families of proteins, RNases. Hence, it was impossible to bases in length between two adult connexins and pannexins, that form gap determine whether the molecules that mesenchymal stem cells. Figure 1B junction channels (Herve et al. 2005). moved from cell to cell were single depicts how a morpholino might gain In invertebrates another family of nucleotides, multimers or combinations access to a gap junction channel. We unrelated proteins, the innexins, serves of both. found that synthetic nucleotides like the same function. Plant cells also those depicted in Fig. 1 up to 24 bases communicate with each other; the Because of the increasing therapeutic in length traversed gap junction structure they employ is called the importance of siRNA, we decided to channels composed of connexin 43, but plasmodesmata. Its pore diameter and reinvestigate the permeability of gap not those composed of connexins 32 length are both two orders of junctions to short strands of and 26. magnitude larger than that of any nucleotides. We began by constructing vertebrate gap junction channel and are tagged strands of nucleotides These studies with morpholino known to allow passage of large solutes (morpholinos) of defined length and oligonucleotides demonstrate that the including siRNAs (Derrick et al. 1992; width. These oligonucleotides were transfer of siRNA from cell to cell via Staehelin, 1997; Vionnet, 2005). resistant to the cells’ nucleases gap junctions might be possible. They (Mudziak et al. 1996). also suggest that the permeability In addition to passing ions to allow could be connexin-specific. To test current flow, gap junction channels To determine experimentally if gap whether siRNA itself could cross gap composed of connexins are permeable junctions are permeable to morpholinos junctions, we needed to identify an to a variety of larger molecules we delivered a known concentration siRNA known to silence a specific including nucleotides such as cAMP through a patch pipette to one cell of a gene. We chose one directed against polymerase β, a gene involved in DNA repair (Polosina et al. 2004). To investigate possible cell-to-cell transfer of the siRNA, three cell populations were transfected with a cDNA encoding polymerase β siRNA. Cell type 1 expressed connexin 43, cell type 2 expressed connexins 32 and 26, while cell type 3 was communication deficient (expressing no connexins). For each cell type we co-cultured wild­ type cells not expressing siRNA with cells transfected to express the siRNA against polymerase β. The experimental protocol then called for separation of the two cell populations to determine if siRNA transfer resulted in a reduction of polymerase β message in the wild­ type cells. We found that cells expressing connexin 43 were unique in transferring the siRNA from the transfected to the wild-type cells, Figure 1. A, The stem cell to stem cell gap junctional permeability to a 12 base morpholino is shown. B, Any rod resulting in knockdown of polymerase shaped oligonucleotide would be free the rotate about its axis in the cytoplasm as indicated by the dashed curved β message. These results confirmed lines. The illustration depicts a 24 base morpholino with dimensions of 1.1 nm by 7.6 nm. To enter the gap junction channel a lesser angle of rotation (solid double arrows) is required. This illustrates that on the basis of the what our studies with the synthetic molecular rotation of a rod shaped species there is a finite probability of entrance into the channel. oligonucleotides had already suggested:

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 17 that gap junction channels composed of connexin 43 are sufficiently permeable to siRNA to facilitate silencing of a gene in an adjoining wild-type cell. The siRNA in this case was 22 bases long with dimensions of 1.1 by 7.4 nm and molecular weight of approximately 5kD.

The permeability of gap junctions to siRNA demonstrates a new function, that of facilitating gene silencing from one cell to another. It suggests the potential for a source cell producing a siRNA, to affect gene expression within a syncytium of cells if an appropriate connexin is expressed. Yet, to understand how effective one cell might be in silencing gene activity in neighbouring cells we need to know how far the siRNA can move within a Figure 2. A, illustrates a linear array of cells linked by gap junctions where one cell is a source of a specific multicellular array. Junctional siRNA, producing that particular species at some rate S (synthesis rate). In all the cells the siRNA is subject to a degradation rate (k). The arrows indicate the transjunctional flux between cell pairs along the array. With distance permeability is one of three important the concentration, C1, … C(x), declines and hence the thickness of the arrow is reduced to signify less flux. B, factors that can limit the spread of illustrates the hypothetical distribution of the siRNA within the array at steady state where the concentration of the siRNA throughout a tissue. The other siRNA is a constant in the source cell and the subsequent concentrations in adjacent cells are subject to two are synthesis and degradation rates degradation without de novo synthesis in these cells. Two cases are shown with high (solid line) and low (dashed line) degradation rates. for the siRNA. Figure 2 is a simplified illustration of the role these factors play in the spread of siRNA within a tissue. (Potapova et al. 2004). These same Kododny GM (1971). Evidence for transfer of macromolecular RNA between mammalian cells in culture. Exp Cell Res 65, 313–324. Of interest is that siRNA from either an cells elicited no demonstrable rejection endogenous or an exogenous source when maintained in canine heart for 6 Mudziak RM, Barofsky E, Barofsky DF, Weller DL, Huang SB & weeks (Plotnikov et al. 2005), Weller DD (1996). Resistance of morpholino phosphorodiamidate survives and functions for many hours oligomers to enzymatic degradation. Antisense Nuc Acid Drug Dev 6, to days (Aliksy & Davidson, 2004) suggesting the possibility of their use 267-272 suggesting that siRNA movement from from allogeneic sources. Plotnikov AN, Shlapakova IN, Szabolcs MJ, Danilo PJ, Lu Z, a source cell to those at distances of Potapova I, Lorell BH, Brink PR, Robinson RB, Cohen IS & Rosen even a number of cell diameters away In summary, the property of siRNA to MR (2005). Adult human mesenchymal stem cells carrying HCN2 gene perform biological pacemaker function with no overt rejection is possible. permeate gap junction channels for 6 weeks in canine heart. Circulation 11, 2II-221. composed of connexins opens a door to siRNA is selective in silencing its gene targeted systemic silencing. How Polosina YY, Rosenquist TA, Grollman AP & Miller H (2004). Knockdown of DNA polymerase by RNA interference: recapitulation target and as such presents an ideal rapidly we avail ourselves of this of null phenotype. DNA Repair 3: 1469 -1474. therapeutic agent to silence the opening and employ cell-based delivery Potapova I, Plotnikov A, Lu Z, Danilo P, Valiunas V, Qu J, Doronin expression of proteins associated with for siRNA-based therapeutics remains S, Zuckerman J, Shlapakova I, Gao J, Pan Z, Herron A, Robinson various disease states in a variety of to be seen. RB, Brink PR, Rosen M & Cohen IS (2004). Human mesenchymal stem cells as a gene delivery system to fabricate a biological substrates. Yet to date its therapeutic pacemaker. Circ Res 94, 952-959. application has been limited by an PR Brink, V Valiunas, inability to deliver the siRNA S Doronin, I Potapova, Staehelin AL (1997). The plant ER: a dynamic organelle composed of a large number of discrete functional domains. Plant J 116, 1151­ selectively to the interiors of the target RB Robinson,* MR Rosen,* 1165. cells. Given our experimental results, RT Mathias, IS Cohen Valiunas V, Doronin S, Valiuniene L, Potapova I, Zuckerman J, the potential for siRNA to be delivered Departments of Physiology and Biophysics at SUNY at Walcott B, Robinson RB, Rosen MR, Brink PR & Cohen IS (2004) by one cell to a syncytium and affect Stony Brook, New York and *Department of Human mesenchymal stem cells make cardiac connexins and form gene expression in many other cells Pharmacology, Columbia University, New York, USA functional gap junctions. J Physiol 555, 617-626 connected by gap junctions raises the References Tsien, R.W & Weingart R (1976). Inotropic effect of cyclic AMP in calf possibility of using siRNA-loaded cells Derrik PM, Barker H, Oparka KJ (1992). Increase in plasmodesmatal ventricular muscle studied by a cut end method. J Physiol 260, 117­ permeability during cell-to-cell spread of tobacco rattle virus from 142. as the platform for a targeted delivery individually inoculated cells. Plant Cell 4, 1405-1412. system. We have recently demonstrated Valiunas V, Polosina Y, Miller H, Potapova I, Valiuniene L, Doronin S, Mathias RT, Robionson RB, Rosen, MR Cohen IS & Brink PR that adult human mesenchymal stem Goldberg G, Valiunas V & Brink PR (2004). Selectivity permeability of gap junction channels. Biochem Biophys Acta 662, 96-101. (2005). Connexin-specific cell to cell transfer of short interfering RNA cells express connexin 43 (Valiunas et by gap junctions. J Physiol 568, 459-468. al. 2004) and can deliver a pacemaker Herve JC, Phelan P, Bruzzone R & White TW (2005). Connexins, innexins and pannexins: bridging the communication gap. Biochem Voinnet O (2005). Non-cell autonomus RNA silencing. FEBS Letters gene to the cardiac syncytium Biophys Acta 1719, 3-5. 579, 5858-5871.

Physiology News | No. 63 | Summer 2006 | www.physoc.org 18 PN FEATURES

Working out aerobic fitness The cardiac and vascular factors that improve endurance performance, aerobic fitness, and exercise capacity are being studied down to subcellular levels in experimental models of exercise training

In the late 1990s, we combined cellular fitness and health (Kemi et al. 2002, cardiology and exercise physiology to Wisloff et al. 2001). Today, the study the biological causes of why guidelines regarding exercise training physical activity and exercise training as a therapeutic approach need are beneficial in both health and refinement and fine-tuning. An disease. Physical activity and exercise important aspect is therefore that training increases function and prevents exercise and the effects on exercise capacity must be controlled, in terms of disease in the cardiovascular system, Clockwise from top right: Ole Kemi, Per Haram, improves the outcome in patients with measuring VO2max accurately and in Oyvind Ellingsen and Ulrik Wisloff manifest disease and improves the terms of how much exercise is prospects of rehabilitation. In fact, performed causing the outcome. An lactate levels (peaking at 6-7 mM) and several sets of data strongly suggest intensity-controlled treadmill model is respiratory exchange ratios above 1.05 that the level of aerobic fitness, as currently the only suitable model when reaching VO2max, strongly suggest determined by metabolic equivalents or allowing this. Custom-made treadmills that the animals were running to maximal oxygen uptake (VO2max), with metabolic chambers for both mice exhaustion, and that VO2max was directly links to health and survival and rats were found to be reproducible assessed validly. Thus, the current prospects, even when traditional risk for measuring oxygen uptake (VO2) procedures for assessing VO2 and factors are present or the patient is on over a range of running intensities. VO2max by large resemble common conventional medical treatment (Myers Also, ramp test protocols for measuring protocols used by applied exercise et al. 2002, O’Neill et al. 2005). It is VO2max were established; highest values physiology laboratories and clinical clear that changes originating from the obtained at or around 25º inclination services working up athletes as well as cell or molecule of the heart and artery indicated that inclined running is patients. For instance, respiratory must accompany and possibly explain necessary to provoke VO2max. Moreover, exchange ratios and blood lactate levels the beneficial effects. a good agreement between heart rate as described above suggest that the and VO2 up to maximal levels during individual, whether it be man or mouse, Thus, we established experimental running to exhaustion; but with is exercising above the aerobic window models to study research questions maximal heart rate occurring after and well into where anaerobic pertaining to biological mechanisms of VO2max, and several-fold elevated blood metabolic processes kick in. These are therefore common criteria used to determine whether or not a ‘true’

VO2max is reached.

Next, we investigated the effects of prolonged high-intensity (85-90% of

VO2max, 5 days/week) interval exercise training on the heart and artery, or the cells that make up those organs (Kemi et al. 2002, 2004, 2005, Wisloff et al.

2001). As VO2max was measured in each animal every week, the running speed was continuously adjusted to maintain the relative training intensity individually. 8-10 weeks of regular

training improves VO2max by ~50-70%; slightly more in rats than mice. Correspondingly, maximal aerobic velocity, i.e. the running velocity where

VO2max was reached, improved similarly. Moreover, the running economy, measured as the cost of

Figure 1. Effects of regular exercise training on VO2max and the cardiomyocyte. oxygen to run at a certain speed, was

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 19 reduced by ~20% during submaximal endothelium. We find that regular weeks, and arterial improvement was running velocities, whereas the exercise training improves endothelial completely lost within 2 weeks of respiratory exchange ratio was reduced function, as the acetylcholine detraining. Thus, exercise-induced by ~5% at those velocities. As for the concentration evokes half-relaxation beneficial adaptations appear heart, right and left ventricular masses decreased ~4-fold in trained animals. vulnerable; what takes months to increased 20-30%, which was There was no difference with an improve, only takes weeks to lose again supported by echocardiography exogenous NO donor (nitroprusside) or if not properly maintained. recordings, whereas left ventricular after blocking NO synthase (L-NAME), cardiomyocytes increased in size by adding further weight to the argument. The rigorous control of exercise ~20%, due to proportional cell length intensity and volume, as well as and width increase. Cell contractile Given that regular exercise training adaptations, in terms of VO2max is function, measured at physiological considerably affects the morphology unprecedented in experimental studies; electrical stimulation frequencies, also and physiology of the cell, we moved with this scheme, exercise training improves after exercise training; the on to study the plasticity of the moves from ‘physical activity’ to an magnitude of shortening increased 30­ changes, by intervening with different entity in which responses may be 40%. In rats, this appears to be largely exercise training programs. We correlated to dose, or the cellular and explained by improved Ca2+ sensitivity, compared the effects of high (85-90% molecular adaptations may be 2+ rather than by more Ca being of VO2max) vs. moderate (65-70%) correlated to changes in VO2max or available to the myofilament intensity exercise, where 1 hour was overall, whole-body function. We machinery, as is the case in mice allocated to exercise per day, 5 conclude that aerobic fitness is closely (unpublished data). Detailed studies of days/week (Kemi et al. 2005). related to cardiomyocyte contractile the myofilament responsiveness to Moderate intensity increased VO2max capacity and acetylcholine-mediated given free [Ca2+] in permeabilized cells only ~half as much as high intensity, endothelial function, and that the support our data that less Ca2+ is needed paralleled by similar reductions in the magnitude of training-induced to evoke the same response after magnitude of changes on cardiac adaptations depends on exercise training in rats (Diffee et al. 2001). myocyte size, fractional shortening, intensity. 2+ Myocytes from the hearts of trained Ca sensitivity, and time-courses of the 1 contraction-relaxation cycle and the Ole Johan Kemi animals also contract and relax more 2 [Ca2+] transient. Thus, the exercise­ Per Magnus Haram quickly than those from untrained 2,3 animals, which coincides with a more induced adaptations in the cardiac Ulrik Wisløff 2,3 rapid increase and decay of the [Ca2+] myocyte depend upon intensity of Øyvind Ellingsen transient. Taken together, the structural exercise. Artery endothelial function 1 had a slightly different dose-response Institute of Biomedical and Life Sciences, University of and functional adaptations of the Glasgow, UK relationship. Acetylcholine-mediated cardiac muscle cell to long-term 2Department of Circulation and Medical Imaging, exercise explains at least partly why NO-dependent vasoreactivity increased Norwegian University of Science and Technology, whole-heart function in trained athletes with exercise, and reached near­ Trondheim, Norway is better than in untrained; increased maximum level of adaptation with 3St Olavs Hospital, Trondheim, Norway stroke volume and cardiac output being moderate exercise, and only a trend for hallmarks of vigorous exercise over higher sensitivity was observed References months and years. The cellular findings between moderate and high intensity. Diffee GM, Seversen EA, Titus MM (2001). Exercise training increases the Ca2+ sensitivity of tension in rat cardiac myocytes. J also provide possible reasoning for Appl Physiol 91, 309-315. better diastolic filling of the ventricles In a different study (Kemi et al. 2004), Kemi OJ, Haram PM, Loennechen JP, Osnes, J-B, Skomedal T, and the concept of athlete’s heart. we investigated the time-course of Wisloff U, Ellingsen O (2005). Moderate vs. high intensity: differential adaptations during training and effects on aerobic fitness, cardiomyocyte contractility, and endothelial function. Cardiovasc Res 67, 161-72. With increased cardiac output, the detraining, that is, how quickly do the artery needs to accordingly secure changes occur when a high-intensity Kemi, OJ, Haram PM, Wisloff U, Ellingsen O (2004). Aerobic fitness exercise program is implemented, and is associated with cardiomyocyte contractile capacity endothelial sufficient conductance of blood to the function in exercise training and detraining. Circulation 109, 2897­ skeletal muscles. One way of doing so how quickly do the effects vanish after 2904. withdrawing regular exercise? Whereas would be increasing the arterial Kemi OJ, Loennechen JP, Wisloff U, Ellingsen O (2002). Intensity­ structural diameter permanently, but an it takes about 8-10 weeks of training controlled treadmill running in mice: cardiac and skeletal muscle hypertrophy. J Appl Physiol 93, 1301-1309. alternative way is to improve the for VO2max, cardiac myocyte size, 2+ vasodilatory capacity. The main contractile function and Ca handling Myers J, Prakash M, Froelicher V, Do D, Partington S, Atwood JE vasorelaxive messenger is nitric oxide to adapt, most of these effects vanish (2002). Exercise capacity and mortality among men referred for exercise testing. N Engl J Med 346, 793-801. (NO), which when released from the very quickly when regular exercise is endothelial cell ultimately reduces stopped. After regular exercise training O'Neill JO, Young JB, Pothier CE, Lauer MS (2005). Peak oxygen 2+ consumption as a predictor of death in patients with heart failure intracellular [Ca ] in the smooth for 10 weeks, half the increase in receiving beta-blockers. Circulation 111, 2313-2318. myocyte, causing relaxation. This VO2max was lost within 2 weeks, most of pathway is stimulated by acetylcholine the effects on cardiomyocyte structure Wisloff U, Helgerud J, Kemi OJ, Ellingsen O. Intensity-controlled treadmill running in rats: VO2max and cardiac hypertrophy. Am J that speeds up NO production in the and function disappeared within 2-4 Physiol Heart Circ Physiol 280, H1301-H1310.

Physiology News | No. 63 | Summer 2006 | www.physoc.org 20 PN FEATURES

Erythropoietin controls ventilation in hypoxia The presence of erythropoietin in blood and brain increases the ventilatory response of mice exposed to oxygen deprivation. Expression of the Epo receptor in the respiratory centre as well as in the carotid bodies suggests that Epo controls the hypoxic ventilatory response at the central (brainstem) and peripheral (carotid body) level

Erythropoietin (Epo) does far more than ‘just’ increasing red blood cell number. Originally discovered as a ‘blood hormone’ and widely applied to anaemic patients, Epo has the potential to become a multipurpose drug. It all began back in 1993, when Ryuzo Sasaki and his Japanese co-workers reported the presence of a functional Epo receptor in rat pheochromocytoma PC12 cells, an established cell line with neuronal characteristics (Sasaki et al. 2001). A few years later Sasaki’s research team, as well as our own, Above, left: Max Gassmann, Omolara Ogunshola and Jorge Soliz; Above, right: demonstrated that Epo and its receptor Max Gassmann with Ryuzo Sasaki; Right: Joseph Vincent are both expressed in the mammalian brain, including human, and that cerebral Epo is expressed in an oxygen­ clinical tests indicated that Epo-treated occurs at high altitude), pulmonary dependent manner. Considering that the patients recovered better and faster ventilation and arterial oxygen content blood-brain-barrier cannot be crossed compared to the saline-treated control are elevated by two complementary by Epo, we predicted a local function patients. systems: the neuronal respiratory of Epo in the brain upon binding to its response (elaborated through the central receptor, but which one(s)? Because Epo has neuroprotective and peripheral nervous system) leading effects, we and others tested the impact to increased minute ventilation thereby Several elegant experiments performed of rhEpo on other neuronal tissues such increasing tissue oxygenation, and the by various teams on different rodent as the spinal cord and the retina renal-derived Epot that activates models of stroke revealed that Epo (reviewed in Gassmann et al. 2003). As erythropoiesis in the bone marrow, thus exerts a protective function upon predicted, we observed that the Epo augmenting the blood’s oxygen experimentally-induced brain ischemia. receptor is also expressed in the retina. carrying capacity. Very recently we As early as 2002, Hannelore Ehrenreich Surprisingly, an increased level of Epo reported for the first time that these two and her team from Göttingen in the retina was beneficial as it systems do crosstalk via Epo (Soliz et (Germany) published the first, and so prevented mice from light-induced al. 2005). far unique, report on Epo's retinal degeneration (Grimm et al. neuroprotective function in man. 2002). Epo’s protective effects are even Carotid bodies are sensory organs Application of recombinant human Epo more widespread, as it has been shown whose stimulation by hypoxia activates (rhEpo) in stroke patients was by different teams that Epo exerts a chemoreflex pathway. The sensory beneficial when given within eight beneficial effects when applied in information is relayed to brainstem hours of the ischemic insult (Ehrenreich several animal models of heart infarct. neurons that in turn modulate et al. 2002). Apart from the tendency to Moreover, we very recently reported compensatory ventilatory adjustments. reduce the infarct volume, several that Epo has protective effects on Carotid bodies are among the most gentamicin-induced auditory hair loss vascularized organs in the body and are (Monge et al. 2006). mainly stimulated by the decline of

PaO2. As PC12 cells mimic carotid Interestingly, all reports on Epo’s non­ body type I cells, we hypothesized that erythropoietic functions point towards a these peripheral chemoreceptors might protective effect of this cytokine upon be also activated by Epo. To address injury (mostly ischemia). We this question we placed the animals into considered that Epo might also exert a a plethysmograph (Fig. 1) and physiological function, and indeed it measured HVR in wild type mice upon does; Epo controls the hypoxic injection of 2000 U/kg rhEpo. Epo­ ventilatory response (HVR) in mice. injected animals showed higher Figure 1. A mouse in a plethysmograph. During physiological hypoxia (as respiratory frequency but lower tidal

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 21 volume than saline-injected controls when exposed to severe hypoxia (6%

O2). These data suggest that Epo has an impact on carotid body cells, most probably by binding to the Epo receptor. We found a dense staining of EpoR in the carotid body, apparently localized within islets of chemosensitive cells. This observation implies that peripheral chemoreceptors can be activated by circulating Epo.

We also suspected that brain-derived Epo modulates ventilation by interacting with brainstem respiratory neurons. To test this hypothesis we used a transgenic mouse line (termed Figure 2. Hypoxic ventilatory response after bilateral transection of the carotid sinus nerve (chemodenervation) in Tg21) that constitutively overexpresses WT and Tg21 mice. The graph shows ventilation (VE) under normoxia (21% O2), mild hypoxia (10% O2) and severe hypoxia (6% O2). Both chemodenervated mouse lines increased ventilation at mild hypoxia due to an human Epo in brain but shows normal increased respiratory rate. However at severe hypoxia, while chemodenerved WT mice showed a very Epo plasma levels (Wiessner et al. pronounced respiratory depression, chemodenervated Tg21 maintained elevated ventilation. WT mice had to be

2001). Tg21 mice showed improved removed from the plethysmographic chamber after 5-10 min of severe hypoxia (6% O2). The closed triangle ventilatory response to severe acute indicates 15 minutes of gradual reduction of FIO2. *p<0.01. Animals per group = 6-7 (from Soliz et al. 2005). hypoxia and improved ventilatory production and lung ventilation to Obviously, Epo will keep us all very acclimatization to chronic hypoxic improve the net oxygen inflow to busy, as the ‘old’ drug has many ‘new’ exposure. Interestingly, following tissues and cells. functions that await detailed bilateral transection of the carotid sinus investigation. nerves that uncouple the brain from the In considering the millennial population carotid body (e.g chemodenervation), of high altitude dwellers, an obvious Jorge Soliz 1 question arises regarding the impact of Tg21 mice adapted their ventilation to Joseph Vincent2 acute severe hypoxia while plasma and cerebral Epo in the Omolara Ogunshola1 chemodenervated wild type (WT) ventilatory adaptation to high altitude. Max Gassmann 1 animals developed a life-threatening Do high altitude inhabitants have 1 apnoea (Fig. 2). higher concentration of Epo in neural Institute of Veterinary Physiology, Vetsuisse Faculty, tissue? Does systemic Epo in high and Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Switzerland These results imply that Epo in the altitude dwellers enhance stimulation of 2Department of Paediatrics, Laval University, Centre de brain modulates ventilation. carotid body functions? In keeping with Recherche, Hopital St-Francois d’Assise, Quebec, Immunohistochemical analysis revealed this, high altitude de-adaptation Canada expression of the Epo receptor in the syndrome is inherent in highlander brainstem’s respiratory centres. populations. In patients suffering from References Additionally, we also provided so-called chronic mountain sickness, Ehrenreich H, Hasselblatt M, Dembowski C, Cepek L, Lewczuk P, evidence that Epo modulates breathing Stiefel M, Rustenbeck HH, Breiter N, Jacob S, Knerlich F, Bohn M, hypoventilation is associated with Poser W, Ruther E, Kochen M, Gefeller O, Gleiter C, Wessel TC, De control by alteration of excessive erythrocytosis, suggesting Ryck M, Itri L, Prange H, Cerami A, Brines M & Siren AL (2002). catecholaminergic metabolism in Erythropoietin therapy for acute stroke is both safe and beneficial. that the connection between ventilation Mol Med 8, 495-505. brainstem. Taken together, these and Epo signals is failing. Gassmann M, Heinicke K, Soliz J & Ogunshola OO (2003). Non­ observations indicate that brain-derived Consequently, the question about the erythroid functions of erythropoietin. Adv Exp Med Biol 543, 323-330. Epo is a key player that modulates implication of neural and plasma Epo Grimm C, Wenzel A, Groszer M, Mayser H, Seeliger M, Samardzija neural respiratory control by acting on in chronic mountain sickness has to be M, Bauer C, Gassmann M & Reme CE (2002). HIF-1-induced the central nervous system. erythropoietin in the hypoxic retina protects against light-induced answered. retinal degeneration. Nat Med 8, 718-724. In summary, we have demonstrated that Finally, Epo is on the doping list Monge A, Nagy I, Bonabi S, Schmid S, Gassmann M & Bodmer D (2006). The effect of erythropoietin on gentamicin-induced auditory high Epo levels in brainstem respiratory because it augments the blood’s oxygen hair cell loss. Laryngoscopy 116, 312-316. neurons improve ventilatory response carrying capacity and thus the oxygen Sasaki R, Masuda S & Nagao M (2001). Pleiotropic functions and and acclimatization to hypoxia. delivery to tissue. Considering that an tissue-specific expression of erythropoietin. News Physiol Sci 16, Furthermore, a link between Epo and increased level of plasma Epo 110-113. respiratory control is provided through stimulates carotid bodies under hypoxic Soliz J, Joseph V, Soulage C, Becskei C, Vogel J, Pequignot JM, Ogunshola O & Gassmann M (2005). Erythropoietin regulates the peripheral nervous system by the conditions, it is tempting to speculate hypoxic ventilation in mice by interacting with brainstem and carotid carotid bodies that express the Epo that Epo-mediated improved bodies. J Physiol 568, 559-571. receptor. In other words, Epo is a performance in athletes is also partially Wiessner C, Allegrini PR, Ekatodramis D, Jewell UR, Stallmach T & due to enhanced ventilatory and/or Gassmann M (2001). Increased cerebral infarct volumes in multifunctional signalling molecule polyglobulic mice overexpressing erythropoietin. J Cereb Blood Flow physiologically acting on red blood cell sympathetic capacity. Metab 21, 857-864.

Physiology News | No. 63 | Summer 2006 | www.physoc.org 22 PN FEATURES

Bursting with currents Vasopressin secretion varies with serum osmolality. We have identified a new osmosensitive current that may alter the firing patterns of the cells that release this important hormone

cell bodies express non-selective stretch afterpotential (DAP) that is activated by inactivated cation channels (SICs) action potentials and that increases the whose activity is determined by likelihood of subsequent action osmotically-evoked changes in cell potentials (Roper et al. 2004). The volume (Oliet & Bourque, 1993). amplitude of the DAP summates during When exposed to hypo-osmolar repetitive firing, leading to the solutions, the MNCs stretch, causing sustained depolarization that underlies the SICs to close. When the external the phasic burst. Ca2+ dependent K+ solution is hyper-osmolar, however, the currents are also activated by action MNCs shrink, which relieves the potentials and these currents may be membrane tension and allows the SICs involved in slowing firing during a to open. This allows cations to flow burst. Although activation of Ca2+ into the cell, causing it to depolarize dependent currents is known to and become more responsive to terminate bursts in other types of excitatory inputs. This process is neurons, they do not appear to play Wenbo Zhang (top) and Thomas Fisher central to explaining how the MNCs such a role in MNCs. Measurements increase their firing rate in response to of Ca2+ levels during a burst suggest When the osmolality of our blood increases in osmolality. that the activation of Ca2+ dependent increases, our magnocellular + neurosecretory cells (MNCs) start to The initiation of a burst depends on a K currents should be maximized well 2+ burst (electrophysiologically speaking, Ca dependent depolarizing before the termination of the phasic that is). These neurones fire few action potentials when our blood is hypo­ osmolar, but progressively increase their rate of firing as the osmolality increases (Bourque & Oliet, 1997). This is part of an important homeostatic mechanism that maintains our body fluid balance because these cells release either vasopressin (VP), which causes the retention of water in the kidneys, or oxytocin (OT), which causes (in some species) excretion of sodium. The release of these two hormones into the circulation is increased as the firing rate of the MNCs is increased. VP­ releasing MNCs, furthermore, adopt a pattern of firing composed of bursts of action potentials lasting tens of seconds alternating with rest periods of roughly equal length. This pattern of firing, known as phasic firing, maximizes VP release.

The electrophysiological mechanisms underlying the osmosensitive changes in firing rate and pattern are incompletely understood. Although Figure 1. Osmosensitive currents and the regulation of MNC firing. The diagram above illustrates typical patterns some of the increase in firing rate is of firing exhibited by VP-ergic MNCs in the presence of different external osmolalities. While the MNCs are due to an increase in excitatory inputs electrically silent in hypo-osmolar solutions, increases in osmolality cause progressive increases in the firing rate onto the MNCs, the MNCs are also and the eventual adoption of a phasic bursting pattern of firing. These changes are associated with an increase in VP secretion. The activation of the SIC causes depolarization of the MNCs leading to an increase in inherently osmosensitive. Patch clamp responsiveness to excitatory inputs and an increase in firing rates. The activation of the OC might be important in experiments have shown that the MNC mediating the transition to burst firing by inhibiting repetitive firing.

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 23 bursts (Roper et al 2004). The inactivation, suggest that it could action potentials. Differential mechanisms underlying the termination increase in amplitude slowly over the activation of the SIC and the OC might of the long phasic bursts of MNCs are course of a burst of action potentials. contribute to a variety of firing patterns, therefore not entirely clear. One The OC might thus be expected to such as slow irregular or fast current model posits that influence the MNC firing rate and/or continuous firing, or firing in short or somatodendritic release of the K-opioid pattern. phasic bursts. peptide dynorphin during a burst acts on the MNCs in an autocrine fashion to The ionic selectivity of the OC has not We have therefore identified an suppress the DAP, thereby leading to yet been established, but our evidence osmosensitive current that is likely to burst termination (Brown & Bourque, suggests that the current may be a K+ be a K+ current and that may be 2004). current. Although the current may be involved in the regulation of MNC carried by Na+ in the absence of K+, the firing in response to changes in the It is also possible, however, that addition of even a small quantity of K+ external osmolality. We are presently osmotic activation of a current or into the internal recording solution trying to identify the current in the currents other than the SIC might be results in block of Na+ flux. When K+ hope of finding specific modulators that involved in the transition to burst firing. is then added to the external solution, would help in identifying its Since MNCs exposed to high voltage steps result in an inward flux of physiological role. It remains to be osmolality may go through a stage of K+. These data suggest that the OC seen whether the OC, like the SIC, is rapid continuous firing before mechanosensitive. Alternatively, the may be a voltage dependent K+ current. converting to a phasic firing pattern current may be modulated by changes Pharmacological experiments support (Bourque & Renaud, 1984), there may in second messenger systems mediated this conclusion. The OC can be be a separate process that is necessary 2+ + by changes in osmolality. Further for the adoption of burst firing. We blocked by either Ba or Cs ions, both characterization of the OC may help to + therefore sought to determine whether of which are known K channel understand the osmosensitive regulation there are any other ion channels blockers. The OC can be activated, of VP and OT release and the modulated by changes in osmolality. however, in the presence of a large transitions between firing patterns As reported in the August issue of The concentration of TEA, suggesting that it observed in the MNCs. + Journal of Physiology, we recently is relatively insensitive to this K identified another current in isolated channel blocker. Acknowledgements MNCs that increases as a function of Work in our laboratory has been + osmolality (Liu et al. 2005). Unlike the What could be the function of a K supported by the Canadian Institutes of SIC, this osmosensitive current (the current that turns on as osmolality is Health Research and the Saskatchewan OC) is also voltage sensitive and is increased? It appears paradoxical that a Health Research Foundation through activated during voltage clamp by steps K+ current would be activated by a the CIHR Regional Partnership to potentials greater than -60 mV. stimulus that causes an increase in the Program by an operating grant and by a Although the amplitude of the OC output from these cells (i.e. the release New Investigator Award to TEF. varies from cell to cell, there is a of VP). The answer to this question marked increase in amplitude may be that the OC has a role in Wenbo Zhang (approximately two-fold) in greater mediating the transition into phasic Thomas E Fisher than 60% of cells tested. The OC is bursting. By slowly activating during Department of Physiology, University of Saskatchewan, therefore the first voltage gated current repetitive firing, the OC might act as a Saskatoon, Canada identified in the MNCs that is sensitive brake that temporarily silences a cell, to osmolality. The slow activation of thereby preparing it to respond to References this current, and the lack of excitatory inputs with another burst of Bourque CW & Oliet SH (1997). Osmoreceptors in the central nervous system. Annu Rev Physiol 59, 601-619.

Bourque CW & Renaud LP (1984). Activity patterns and The Journal of Physiology osmosensitivity of rat supraoptic neurones in perfused hypothalamic explants. J Physiol (Lond) 349, 631-642. Brian Robertson (University of Leeds) has been elected as Deputy Editor-in- Brown CH & Bourque CW (2004). Autocrine feedback inhibition of Chief of The Journal of Physiology to replace Prem Kumar in July. Brian is plateau potentials terminates phasic bursts in magnocellular also The Journal’s Topical Reviews Editor. Gordon Drummond (Royal neurosecretory cells of the rat supraoptic nucleus. J Physiol 557, Infirmary, Edinburgh) will serve as an additional representative of the Editorial 949-960. Board on The Journal of Physiology’s Executive Committee. Liu XH, Zhang W & Fisher TE (2005). A novel osmosensitive voltage gated cation current in Rat Supraoptic Neurones. J Physiol 568, 61­ 68. Other Editors currently serving on The Journal’s Executive Committee are Oliet SH & Bourque CW (1993). Mechanosensitive channels William Large (Editor-in-Chief), Steven Mifflin (International Editor), George transduce osmosensitivity in supraoptic neurons. Nature 364, 341­ Augustine (Designated Senior Editor), Hugh Matthews (Designated Senior 343. Editor Elect), Peter Ellaway and Chris McBain. The Society is represented by Roper P, Callaway J & Armstrong W (2004). Burst initiation and the Chairman of its Executive Committee, the Treasurer and the Chief termination in phasic vasopressin cells of the rat supraoptic nucleus: a combined mathematical, electrical, and calcium fluorescence study. Executive. J Neurosci 24, 4818-4831.

Physiology News | No. 63 | Summer 2006 | www.physoc.org 24 PN FEATURES

Developing insights into Purkinje cell maturation Recent evidence reveals a temporal correspondence between the maturation of spike output properties and cell structure during postnatal Purkinje cell development, which further parallels the development of anatomical, synaptic and behavioural indices of cerebellar maturation

The cerebellum coordinates fine motor immature, synaptic innervation is movements, underlies the adaptation of rudimentary, and the repertoire of ocular responses, and supports the cerebellar-dependent behaviours is learning of some conditioned limited (Altman & Bayer, 1997). By behaviours (Ito, 1984). However, in the third to fourth postnatal weeks the the rat and mouse, these behaviours, anatomy, synaptic connectivity and and the anatomy and physiology that neuronal morphology, as well as Bruce McKay (left) and Ray Turner enable these functions, are either absent behavioural indices of cerebellar in the altricial neonate, or present in an function, all reach adult levels (Altman To this end we studied the structural immature form (Altman & Bayer, & Bayer, 1997). The development of and functional development of Purkinje 1997). these characteristics is known with cells, a cell type of central importance precise temporal resolution. However, to overall cerebellar function, from P0 At birth the cerebellum is limited to a the development of intrinsic neuronal through P90 using whole-cell patch pair of small masses overlying the properties, such as spike output clamp recordings and Neurobiotin cell fourth ventricle, with Purkinje cells patterns, and any relationship to the fills in the in vitro slice preparation arranged in clusters and granule cells development of cell morphology, had (McKay & Turner, 2005). We found present in an external layer of the not been determined with high temporal that both morphological and cortex. Neuronal morphology is resolution for any cerebellar cell type. electrophysiological characteristics develop with a sigmoidal time course: an initial stable immature stage of minimal change from P0 to ~P9, a transition stage from >P9 to ~P18 wherein the conversion from immature to adult characteristics occurs, and finally a stable adult stage >P18 where only minor refinements are noted (McKay & Turner, 2005). For instance, during the stable immature stage Purkinje cells possess a peri-somatic dendritic configuration (e.g., P3 and P9, Fig. 1A). During the transition stage Purkinje cells retract their peri-somatic dendrites and then generate a single primary stem dendrite that rapidly increases in surface area to nearly adult levels (e.g., P12 and P18, Fig. 1A). During the stable adult stage only small morphological changes are observed (Fig. 1A and 1E).

Occurring alongside morphological changes is a parallel shift in the patterns of evoked and spontaneous spike output. Throughout the stable immature stage intracellular stimulation evokes a transient low threshold burst discharge of action potentials (Fig. 1B). However, during the transition stage Purkinje cells proceed through an Figure 1. Development of Purkinje cell structure and function. A, Purkinje cell morphology from P3 to P90. B-D, under-developed form of repetitive high evoked spike output for an immature (P9), intermediate (P15) and adult (P60) Purkinje cell. E, Superimposed Boltzmann fits for the development of Purkinje cell dendrite area and percentage of cells generating the trimodal threshold bursts (Fig. 1C), which pattern of spontaneous spike output. All panels adapted from McKay & Turner (2005). ultimately matures to a stable adult

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES/IMAGES OF MUNICH PN 25 pattern of high threshold burst output cerebellar-dependent behaviours, Joint International Meeting of (Fig. 1D). Following a similar time including pelvis elevation, maintaining course, the percentage of cells balance on a narrow plank, mid air the German Physiological generating the trimodal (tonic, burst, righting, and rearing without support, Society and the Federation of quiet) pattern of spontaneous output is are essentially absent during the first European Physiological 0% throughout the stable immature postnatal week, improve significantly Societies (FEPS) stage and quickly increases to nearly during the second and third postnatal Munich, Germany 100% by the end of the transition stage weeks, and stabilize at adult values by 26–29 March 2006 (Fig. 1E). Co-plotting the Boltzmann the fourth postnatal week, further fits (see McKay & Turner, 2005) of the mirroring the development of Purkinje developmental changes in dendritic cell characteristics (Altman & Bayer, surface area and the proportion of 1997). Purkinje cells generating the trimodal pattern reveals the relative postnatal Thus despite the diversity of these time courses for these two variables. anatomical, synaptic, cellular and This strategy indicates that dendritic behavioural indices, the temporal development precedes the development progression towards adult-like character of trimodal output by a few days, but follows a similar sigmoidal time course importantly reveals that, once initiated, – each characteristic begins with an these characteristics mature at similar initial stage of stable immaturity rates. persisting for approximately the first postnatal week, rapidly progresses Interestingly, the time courses of through a transition stage over the multiple aspects of cerebellar course of only approximately one week, development, including the maturation and then persists in a stable adult stage. of gross anatomy, synaptic innervation and behavioural indices appear to The use of Boltzmann functions to parallel the time course of development analyze a range of developmental we describe for Purkinje cell structure variables may prove useful in and spike output. establishing a comprehensive and quantitative picture of multiple aspects For instance, one measure of gross of postnatal cerebellar development. anatomy – the cross-sectional area of the vermis – changes minimally during Acknowledgements approximately the first postnatal week This work was funded by the Canadian (stable immature stage), undergoes a Institutes for Health Research (RWT, massive expansion to adult size over BEM), the Alberta Heritage Foundation the next two weeks (transition stage), for Medical Research (RWT), the and then plateaus in area (stable adult Killam Trust (BEM) and a Steinhauer stage) (Altman & Bayer, 1997). Also Doctoral Award (BEM). following a sigmoidal time course is the development of synaptic inputs to Bruce E McKay Purkinje cells. Throughout the first postnatal week only a small number of Ray W Turner Purkinje cell parallel fibre and basket Hotchkiss Brain Institute, University of Calgary, Calgary, Alberta, Canada cell synapses are detected, whereas a marked proliferation is noted during the second postnatal week with the References Altman J & Bayer SA (1997). Development of the Cerebellar System. attainment of stable adult levels by the CRC Press, New York. third postnatal week (Larramendi, 1969). Further, the number of climbing Ito M (1984). The cerebellum and neural control. Raven, New York. fibres that synapse onto each Purkinje Larramendi LMH (1969). Analysis of synaptogenesis in the cell is stable at 3-4 during the first cerebellum of the mouse. In Neurobiology of cerebellar evolution and development. ed. Llinas R. American Medical Association, Chicago. postnatal week, decreases to one From the top: Bavarian Evening; Ludwig- Maximilians-Universität; visitors to the Blackwell climbing fibre per Purkinje cell during McKay BE & Turner RW (2005). Physiological and morphological development of the rat cerebellar Purkinje cell. J Physiol 567, 829­ Publishing/Journal of Physiology stand – Lutz the second postnatal week, and remains 850. Pott (former Journal Editor) and Bernhard stable at this level throughout Scelfo B & Strata P (2005). Correlation between multiple climbing Becker from the local organising committee. adulthood (Scelfo & Strata, 2005). fibre regression and parallel fibre response development in the (photos by Mary Wilson and Sue Ecob who Finally, the maturation of known postnatal mouse cerebellum. Eur J Neurosci 21, 971-978. represented The Journal of Physiology).

Physiology News | No. 63 | Summer 2006 | www.physoc.org 26 PN FEATURES

Muscle mass at the top: a likely role for fibre hyperplasia in humans The extreme increase in muscle mass observed after heavy resistance training cannot be simply explained on the basis of muscle fibre hypertrophy, as is commonly assumed

The foremost factor shaping muscle increase of fibres size, may be the phenotype through the quantitative activation of protein synthesis mechanisms is exercise resistance pathways. The expression of Insulin­ training (Fig. 1). Resistance training like Growth Factor-I (IGF-I), induced has been extensively studied and is well by muscle overload, and insulin have known to stimulate muscle hypertrophy. been demonstrated to be responsible for regulation of protein synthesis by Giuseppe D’Antona Interestingly, the level of muscle mass stimulating the Phosphoinositide 3’­ hypertrophy described in various works Kinase/Protein Kinase (PI3K/Akt) appears to depend on the experimental pathway, which in turn results in the The increase in skeletal muscle mass in design of the study. In fact downstream activation of targets response to several stimuli including comparative studies of different required for protein synthesis. Recently intense physical exercise is considered subjects populations (cross-sectional it has also been demonstrated that an obvious example of cell plasticity. studies) have shown higher but activation of Akt is able to induce an The accepted mechanism underlying comparable degree of muscle increase in muscle mass through a such changes is quantitative hypertrophy (MacDougall et al. 1982; dramatic increase in fibre size of modifications of gene expression. D'Antona et al. 2006), whereas individual fibres (Glass, 2005). longitudinal studies of shorter (2-14 In skeletal muscle each gene can be weeks) duration have shown much Notwithstanding an undoubted role of upregulated or downregulated in lower degree of hypertrophy (Aagaard fibres hypertrophy, the traditionally response to several factors including et al. 2001). accepted quantitative mechanism of mechanical load (exercise), neural muscle plasticity is not fully able to discharge and hormones. Gene It is generally assumed that the increase explain the observed changes in muscle products can be quantitatively modified in muscle mass following resistance mass due to resistance exercise. In and new functional and structural training can be fully accounted for by fact, studies performed on resistance features can appear at macroscopic individual muscle fibre hypertrophy athletes have shown a wide range of level. In particular, the ‘quantitative and hypertrophic response appears to muscle hypertrophy without a strict mechanism’ of muscle plasticity seems be fibre-type specific. Whereas correlation with changes in fibre size. to be the major accepted factor by hypertrophy clearly spares slow fibres, This observation is most clear when which skeletal muscle can adapt to it selectively involves fast fibres considering the few available studies on variable functional requirements (Aagaard et al. 2001; D'Antona et al. body builders. These reported only through changes in mass and fibre size 2006). The cause of such fibre-type limited hypertrophy of muscle fibres, (hypertrophy). Quantitative changes of selectivity is unclear and requires future failing to account for the obvious and fibre size are also commonly investigations. extreme hypertrophy of whole muscles considered as the major determinants of (MacDougall et al. 1982; Tesch & load-induced changes in muscle force The mechanism by which heavy work Larsson, 1982). generation. increases muscle mass, through the The observed discrepancy between the mean fibre cross sectional area and the anatomical cross sectional area (CSAanat, cross sectional area of a muscle determined in a plane axial to the long axis of a muscle) of the muscle determined by MRI clearly suggests that hypertrophy cannot fully explain the observed hypertrophy of whole muscle (MacDougall et al. 1982; Tesch & Larsson, 1982).

Which mechanisms underlie such discrepancy? Which other phenomena than fibres Figure 1. Coronal (A, B) and sagittal (C) magnetic resonance images (MRI) of the thigh of a normally active hypertrophy contribute to the striking subject (A) and a body builder (B, C) highlight dramatic increase of skeletal muscle mass following long term heavy resistance training (modified from D’Antona et al. 2006). increase in muscle mass due to long-

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 27

the vastus lateralis of elite body arise in interstitial tissue between builders and untrained subjects. By existing fibres (Fig. 2). applying these values to a simple theoretical model of muscle Regarding the possible signals involved architecture, we excluded the idea that in activation of quiescent satellite cells, such differences could be entirely a number of extracellular factors are Table 1. Physiological cross sectional area (CSAp) of attributed to differences in pennation known to be involved in muscle the vastus lateralis of a body builder and a control angle between the two groups of regeneration that is triggered in normally active subject (D’Antona G unpublished subjects (D'Antona et al. 2006). response to muscle injury. Some of observation). them (Myogenic Transcription Factors, term and very high intense resistance These results combined with the MRFs, IGF; Fibroblast Growth Factors, training in humans? observation of signs of muscle FGF; Hepatocyte Growth Factor, HGF; regeneration (the appearance of a low Sonic hedgehog, Shh and others) are In a recent study we aimed to help percentage of neonatal myosin involved in the activation of adult stem answer this question by investigating if isoforms) allowed us to hypothesize cells and in their proliferation, while changes in muscle architecture may that hyperplasia represented the most others (MRFs, IGF, Neuregulin, NRG, explain the difference between mean feasible mechanism, overlapping with Shh, Wingless-Int, Wnt) promote fibre area and CSAanat observed in the increase in fibres size, leading to muscle differentiation. Their possible elite male body builders. the observed increase in muscle mass role in determining the hyperplastic after chronic heavy resistance training. response after chronic heavy resistance It is known that the relation between training deserves further investigation. the physiological CSA (CSAphysiol, Unfortunately, to date, only indirect cross sectional area of a individual observations like ours support this Acknowledgments muscle fibres of a muscle, Table 1), and hypothesis in humans. In animal I am grateful to Marco Narici and the anatomical CSA (CSAanat) is not models (rat or chicken) some evidence Francesca Grisù Caliaro for useful constant. The latter relation varies does suggest that hyperplasia can occur discussions on muscle architecture and according to the pennation angle, which in overloaded conditions or after regeneration. This work has been is the angle between the axis of the stretching. supported by a grant from Italian Space fibres and the axis of the muscle Agency (ASI) and World Anti-Doping (Narici et al. 1996). Physical exercise The possible mechanisms involved in Agency (WADA). is known to induce significant exercise-induced muscle hyperplasia Giuseppe D’Antona adjustments of the spatial orientation of are only partially known. It has been the muscle fibres. This varies the demonstrated that muscle overload Department of Experimental Medicine, Human Physiology Unit and Interuniversity Institute of Myology pennation angle, which has been found induces new fibre formation resulting (IIM), University of Pavia, Italy to increase following resistance training from satellite cell activation, and to be larger in body builders than in proliferation, differentiation and fusion, References similar to what happens during controls (Kawakami et al. 1995). An Aagaard P, Andersen JL, Dyhre-Poulsen P, Leffers AM, Wagner A, increase in the pennation angle has thus development (Tamaki et al. 1997). It is Magnusson SP, Halkjaer-Kristensen J & Simonsen EB (2001). A unclear to what extent satellite cell mechanism for increased contractile strength of human pennate been considered as a possible source of muscle in response to strength training: changes in muscle the discrepancy between CSAphysiol proliferation and fusion arise within the architecture. J Physiol 534, 613-623. exercised muscle and the fibre injury and CSAanat. D'Antona G, Lanfranconi F, Pellegrino MA, Brocca L, Adami R, Rossi appears the origin of the regenerative R, Moro G, Miotti D, Canepari M & Bottinelli R (2006). Skeletal In our work, we considered known response. In both rats and chickens muscle hypertrophy and structure and function of skeletal muscle fibres in male body builders. J Physiol 570, 611-627. values of pennation angle and measured subjected to muscle overload there is values of CSAphysiol and CSAanat of evidence that cells formed de novo may Glass DJ (2005). Skeletal muscle hypertrophy and atrophy signaling pathways. Int J Biochem Cell Biol 37, 1974-1984.

Kawakami Y, Abe T, Kuno SY & Fukunaga T (1995). Training­ induced changes in muscle architecture and specific tension. Eur J Appl Physiol Occup Physiol 72, 37-43.

MacDougall JD, Sale DG, Elder GC & Sutton JR (1982). Muscle ultrastructural characteristics of elite powerlifters and bodybuilders. Eur J Appl Physiol Occup Physiol 48, 117-126.

Narici MV, Binzoni T, Hiltbrand E, Fasel J, Terrier F & Cerretelli P (1996). In vivo human gastrocnemius architecture with changing joint angle at rest and during graded isometric contraction. J Physiol 496, 287-297.

Tamaki T, Akatsuka A, Tokunaga M, Ishige K, Uchiyama S & Shiraishi T (1997). Morphological and biochemical evidence of muscle hyperplasia following weight-lifting exercise in rats. Am J Physiol 273, C246-256. Figure 2. Cross sections of rat plantaris muscle after resistance exercise training demonstrate the appearance of degenerating cells (arrowheads in Panel B) as well as regenerating cells at the periphery of normal fibres (arrows Tesch PA & Larsson L (1982). Muscle hypertrophy in bodybuilders. in Panel C). (with permission from Tamaki T et al. 1997). Eur J Appl Physiol Occup Physiol 49, 301-306.

Physiology News | No. 63 | Summer 2006 | www.physoc.org 26 PN FEATURES

Muscle mass at the top: a likely role for fibre hyperplasia in humans The extreme increase in muscle mass observed after heavy resistance training cannot be simply explained on the basis of muscle fibre hypertrophy, as is commonly assumed

The foremost factor shaping muscle increase of fibres size, may be the phenotype through the quantitative activation of protein synthesis mechanisms is exercise resistance pathways. The expression of Insulin­ training (Fig. 1). Resistance training like Growth Factor-I (IGF-I), induced has been extensively studied and is well by muscle overload, and insulin have known to stimulate muscle hypertrophy. been demonstrated to be responsible for regulation of protein synthesis by Giuseppe D’Antona Interestingly, the level of muscle mass stimulating the Phosphoinositide 3’­ hypertrophy described in various works Kinase/Protein Kinase (PI3K/Akt) appears to depend on the experimental pathway, which in turn results in the The increase in skeletal muscle mass in design of the study. In fact downstream activation of targets response to several stimuli including comparative studies of different required for protein synthesis. Recently intense physical exercise is considered subjects populations (cross-sectional it has also been demonstrated that an obvious example of cell plasticity. studies) have shown higher but activation of Akt is able to induce an The accepted mechanism underlying comparable degree of muscle increase in muscle mass through a such changes is quantitative hypertrophy (MacDougall et al. 1982; dramatic increase in fibre size of modifications of gene expression. D'Antona et al. 2006), whereas individual fibres (Glass, 2005). longitudinal studies of shorter (2-14 In skeletal muscle each gene can be weeks) duration have shown much Notwithstanding an undoubted role of upregulated or downregulated in lower degree of hypertrophy (Aagaard fibres hypertrophy, the traditionally response to several factors including et al. 2001). accepted quantitative mechanism of mechanical load (exercise), neural muscle plasticity is not fully able to discharge and hormones. Gene It is generally assumed that the increase explain the observed changes in muscle products can be quantitatively modified in muscle mass following resistance mass due to resistance exercise. In and new functional and structural training can be fully accounted for by fact, studies performed on resistance features can appear at macroscopic individual muscle fibre hypertrophy athletes have shown a wide range of level. In particular, the ‘quantitative and hypertrophic response appears to muscle hypertrophy without a strict mechanism’ of muscle plasticity seems be fibre-type specific. Whereas correlation with changes in fibre size. to be the major accepted factor by hypertrophy clearly spares slow fibres, This observation is most clear when which skeletal muscle can adapt to it selectively involves fast fibres considering the few available studies on variable functional requirements (Aagaard et al. 2001; D'Antona et al. body builders. These reported only through changes in mass and fibre size 2006). The cause of such fibre-type limited hypertrophy of muscle fibres, (hypertrophy). Quantitative changes of selectivity is unclear and requires future failing to account for the obvious and fibre size are also commonly investigations. extreme hypertrophy of whole muscles considered as the major determinants of (MacDougall et al. 1982; Tesch & load-induced changes in muscle force The mechanism by which heavy work Larsson, 1982). generation. increases muscle mass, through the The observed discrepancy between the mean fibre cross sectional area and the anatomical cross sectional area (CSAanat, cross sectional area of a muscle determined in a plane axial to the long axis of a muscle) of the muscle determined by MRI clearly suggests that hypertrophy cannot fully explain the observed hypertrophy of whole muscle (MacDougall et al. 1982; Tesch & Larsson, 1982).

Which mechanisms underlie such discrepancy? Which other phenomena than fibres Figure 1. Coronal (A, B) and sagittal (C) magnetic resonance images (MRI) of the thigh of a normally active hypertrophy contribute to the striking subject (A) and a body builder (B, C) highlight dramatic increase of skeletal muscle mass following long term heavy resistance training (modified from D’Antona et al. 2006). increase in muscle mass due to long-

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 27

the vastus lateralis of elite body arise in interstitial tissue between builders and untrained subjects. By existing fibres (Fig. 2). applying these values to a simple theoretical model of muscle Regarding the possible signals involved architecture, we excluded the idea that in activation of quiescent satellite cells, such differences could be entirely a number of extracellular factors are Table 1. Physiological cross sectional area (CSAp) of attributed to differences in pennation known to be involved in muscle the vastus lateralis of a body builder and a control angle between the two groups of regeneration that is triggered in normally active subject (D’Antona G unpublished subjects (D'Antona et al. 2006). response to muscle injury. Some of observation). them (Myogenic Transcription Factors, term and very high intense resistance These results combined with the MRFs, IGF; Fibroblast Growth Factors, training in humans? observation of signs of muscle FGF; Hepatocyte Growth Factor, HGF; regeneration (the appearance of a low Sonic hedgehog, Shh and others) are In a recent study we aimed to help percentage of neonatal myosin involved in the activation of adult stem answer this question by investigating if isoforms) allowed us to hypothesize cells and in their proliferation, while changes in muscle architecture may that hyperplasia represented the most others (MRFs, IGF, Neuregulin, NRG, explain the difference between mean feasible mechanism, overlapping with Shh, Wingless-Int, Wnt) promote fibre area and CSAanat observed in the increase in fibres size, leading to muscle differentiation. Their possible elite male body builders. the observed increase in muscle mass role in determining the hyperplastic after chronic heavy resistance training. response after chronic heavy resistance It is known that the relation between training deserves further investigation. the physiological CSA (CSAphysiol, Unfortunately, to date, only indirect cross sectional area of a individual observations like ours support this Acknowledgments muscle fibres of a muscle, Table 1), and hypothesis in humans. In animal I am grateful to Marco Narici and the anatomical CSA (CSAanat) is not models (rat or chicken) some evidence Francesca Grisù Caliaro for useful constant. The latter relation varies does suggest that hyperplasia can occur discussions on muscle architecture and according to the pennation angle, which in overloaded conditions or after regeneration. This work has been is the angle between the axis of the stretching. supported by a grant from Italian Space fibres and the axis of the muscle Agency (ASI) and World Anti-Doping (Narici et al. 1996). Physical exercise The possible mechanisms involved in Agency (WADA). is known to induce significant exercise-induced muscle hyperplasia Giuseppe D’Antona adjustments of the spatial orientation of are only partially known. It has been the muscle fibres. This varies the demonstrated that muscle overload Department of Experimental Medicine, Human Physiology Unit and Interuniversity Institute of Myology pennation angle, which has been found induces new fibre formation resulting (IIM), University of Pavia, Italy to increase following resistance training from satellite cell activation, and to be larger in body builders than in proliferation, differentiation and fusion, References similar to what happens during controls (Kawakami et al. 1995). An Aagaard P, Andersen JL, Dyhre-Poulsen P, Leffers AM, Wagner A, increase in the pennation angle has thus development (Tamaki et al. 1997). It is Magnusson SP, Halkjaer-Kristensen J & Simonsen EB (2001). A unclear to what extent satellite cell mechanism for increased contractile strength of human pennate been considered as a possible source of muscle in response to strength training: changes in muscle the discrepancy between CSAphysiol proliferation and fusion arise within the architecture. J Physiol 534, 613-623. exercised muscle and the fibre injury and CSAanat. D'Antona G, Lanfranconi F, Pellegrino MA, Brocca L, Adami R, Rossi appears the origin of the regenerative R, Moro G, Miotti D, Canepari M & Bottinelli R (2006). Skeletal In our work, we considered known response. In both rats and chickens muscle hypertrophy and structure and function of skeletal muscle fibres in male body builders. J Physiol 570, 611-627. values of pennation angle and measured subjected to muscle overload there is values of CSAphysiol and CSAanat of evidence that cells formed de novo may Glass DJ (2005). Skeletal muscle hypertrophy and atrophy signaling pathways. Int J Biochem Cell Biol 37, 1974-1984.

Kawakami Y, Abe T, Kuno SY & Fukunaga T (1995). Training­ induced changes in muscle architecture and specific tension. Eur J Appl Physiol Occup Physiol 72, 37-43.

MacDougall JD, Sale DG, Elder GC & Sutton JR (1982). Muscle ultrastructural characteristics of elite powerlifters and bodybuilders. Eur J Appl Physiol Occup Physiol 48, 117-126.

Narici MV, Binzoni T, Hiltbrand E, Fasel J, Terrier F & Cerretelli P (1996). In vivo human gastrocnemius architecture with changing joint angle at rest and during graded isometric contraction. J Physiol 496, 287-297.

Tamaki T, Akatsuka A, Tokunaga M, Ishige K, Uchiyama S & Shiraishi T (1997). Morphological and biochemical evidence of muscle hyperplasia following weight-lifting exercise in rats. Am J Physiol 273, C246-256. Figure 2. Cross sections of rat plantaris muscle after resistance exercise training demonstrate the appearance of degenerating cells (arrowheads in panel B) as well as regenerating cells at the periphery of normal fibres (arrows Tesch PA & Larsson L (1982). Muscle hypertrophy in bodybuilders. in panel C). (with permission from Tamaki T et al. 1997). Eur J Appl Physiol Occup Physiol 49, 301-306.

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Cardiac channelopathies studied with the dynamic action potential clamp technique

The congenital long-QT syndrome Since the discovery of genetic linkage (LQTS) is an inherited disorder that between cardiac ion channels and the prolongs repolarisation of the LQTS in the mid 1990s, remarkable ventricular myocyte. It often leads to results were obtained in understanding sudden death from cardiac arrhythmias various types of this syndrome. At Géza Berecki (left) and Antoni van Ginneken arising from oscillations during the present, most effort has been devoted to action potential (AP) plateau, documenting the effects of identified despite advances in mathematical specifically torsades de pointes and mutations on the densities and kinetics modeling, the mechanism by which a ventricular fibrillation. It is a of various cardiac ion channels upon given genetic defect leads to the characteristic feature of the cardiac cell heterologous expression. Subsequently, clinically observed electrical disease membrane that even small changes in the consequences of channelopathies often remains obscure: genetic individual ionic currents can shift the for cardiac function are inferred by heterogeneity in LQTS represents a delicate balance between inward and testing the functional effects of the major challenge for modelers as various outward current flow during the plateau experimentally observed changes with mutations in the same gene may use of mathematical models of cardiac and repolarisation and thereby prolong produce different, often inconsistent cells (Wehrens et al. 2002). However, or shorten the AP. functional effects.

The recently introduced ‘dynamic action potential clamp’ (dAPC) technique (Berecki et al. 2005; 2006) represents an alternative approach to investigate LQTS without making assumptions with regard to (altered) kinetic properties of the studied channel(s). As an extension of the dynamic clamp methodology (briefly reviewed by Wilders, 2005), allows the study of various types of LQTSs (e.g. LQT2 and LQT3, which are linked to mutations in the human ether-a-go-go­ related gene, HERG, and sodium channel gene, SCN5A, respectively).

The concept of dAPC is that a selected native ionic current of a ventricular myocyte is effectively replaced with wild-type or mutant current recorded from a transiently transfected human embryonic kidney (HEK)-293 cell. During the dAPC experiment, the ventricular cell and the HEK-293 cell are electrically coupled by means of an electrical circuit. The ventricular cell is in ‘current clamp’ mode on one patch­ clamp setup, whereas the HEK293 cell Figure 1. Implementing cardiac sodium current (INa) in dAPC experiments. A, Overall experimental design. INa is in ‘voltage clamp’ mode on another from the HEK cell is continuously applied to the ventricular cell as an external current input, partly or entirely setup. The command potential for the replacing INa in the ventricular cell. B, “Model cell mode” dAPC with WT (left) or Y1795C INa (right). From top to bottom: subepicardial cell APs elicited at 1 Hz before and after a 2-s pause. APs with mutant INa are prolonged HEK293 cell is the Vm of the compared to WT. Boxed APs from top traces and associated INa on expanded time scale. Note that late INa­ ventricular cell (‘action potential increase after the pause is more pronounced with the mutant (asterisk). Dotted line indicate persistent (late) clamp’), and the current input applied current amplitude, characteristic in type 3 LQTS; peak INa is off scale (arrows); Relationships between the APs and to the ventricular cell is the current selected membrane current components of the human ventricular model cell, showing the changes in the time + + recorded from the transfected HEK293 course of transient outward K current (Ito), slowly and rapidly activating components of the delayed rectifier K 2+ cell, a connection resulting in dAPC current (IKs and IKr, respectively), and L-type Ca current (ICa), along with changes in mutant INa. Adapted from Berecki et al. (2006) with permission from Blackwell Publishing Ltd. condition.

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and subepicardial ventricular cells by adjusting selected membrane ionic currents in the ventricular model cell. Equally, the technically more difficult real cell mode reveals AP waveforms and ion channel kinetics that can be considered close-to-physiological.

The dAPC approach allows rapid and unambiguous determination of the effect(s) of ion channel mutation on the ventricular AP morphology. During experiments, the (altered) shape of the AP directly reflects the effect of a mutation; the frequency dependence of the AP durations, the consequence of a pause on AP morphology as well the arrhythmogenic nature of LQT­ associated mutations can be determined, and special kinetic features of cardiac channels can be revealed. With adequate scaling adjustments and procedures to reduce unwanted and contaminating background currents, this novel technique allows several cardiac ion channels to be studied.

The dAPC technique represents a promising new tool to study various cardiac ion channels and may also prove useful in related fields of research, e.g. in neurophysiology.

Acknowledgement This study was supported by Netherlands Heart Foundation Grant 2001B155.

Géza Berecki Antoni C G van Ginneken Department of Experimental Cardiology, Academic Medical Center, University of Amsterdam, The Netherlands.

References

Figure 2. ‘Real cell mode’ dAPC experiment with WT and mutant HERG current (IHERG). A, Diagram of the dAPC Berecki G, Zegers JG, Verkerk AO, Bhuiyan ZA, de Jonge B, technique that is used to effectively replace the pharmacologically blocked native I of a ventricular cell with I Veldkamp MW, Wilders R & van Ginneken AC (2005). HERG Kr HERG channel (dys)function revealed by dynamic action potential clamp from a HEK293 cell. B, APs from a myocyte and associated WT (top) or R56Q IHERG (bottom), at different technique. Biophys J 88, 566-578. frequencies. The myocyte was successively coupled to HEK-293 cells transfected with WT or R56Q HERG channels. Note the different IHERG waveforms (*, transient IHERG; arrow: sustained IHERG) and frequency dependent Berecki G, Zegers JG, Bhuiyan ZA, Verkerk AO, Wilders R, & van AP prolongation with R56Q. Adapted from Berecki et al. (2005) with the permission of the Biophysical Society Ginneken AC (2006). Long-QT syndrome-related sodium channel mutations probed by the dynamic action potential clamp technique. J (http://www.biophysj.org). Physiol 570, 237-250.

Both a computed model of the human first offers an outstanding Priebe L & Beuckelmann DJ (1998). Simulation study of cellular ventricular cell (Priebe & reproducibility of the results, because electric properties in heart failure. Circ Res 82, 1206-1223. Beuckelmann, 1998) (Fig. 1) or a the implemented (input) wild-type Wehrens XH, Vos MA, Doevendans PA & Wellens HJ (2002). Novel freshly isolated (real) myocyte (Fig. 2) (WT) or mutant current is the only insights in the congenital long QT syndrome. Ann Intern Med 137, can effectively be used in dAPC variable during experimentation. 981-992. experiments, defining the ‘model cell’ Moreover, it allows ‘generation’ of Wilders R (2005). 'Dynamic clamp' in cardiac electrophysiology. J and ‘real cell’ modes, respectively. The subendocardial, midmyocardial (M), Physiol 566, 641

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Reduced mitochondrial efficiency: dysfunction or defence in ageing muscle? Mitochondrial oxidative phosphorylation has been found to become less efficient with age in both mouse and human skeletal muscle. Here, David Marcinek explores mechanisms that can lead to either a protective or pathological effect of this uncoupling

average about 50% fewer ATP mitochondria isolated from aged rodent

molecules per O2 molecules consumed muscle (Harper et al. 2004). Such (reduced P/O) than those in young proton leak may occur as a result of muscle (Fig. 1). This mitochondrial non-specific leak across the IMM uncoupling is consistent with the resulting from damage to membrane reduced in vivo phosphorylation components in aged muscle, or the leak capacity per mitochondrial volume could be mediated through activities of specific membrane proteins, such as the David Marcinek found in the quadriceps from elderly humans (Conley et al. 2000). uncoupling proteins (UCP). These leak Mitochondria are primary sites for Mitochondrial ATP synthesis is coupled pathways are not necessarily mutually aerobic ATP synthesis and generation of to oxygen consumption in the exclusive, but identifying the roles of reactive oxygen species (ROS). They mitochondria through the membrane membrane damage versus UCP also play an important role in potential generated by proton pumping function may be critical to determining regulating cell survival. These multiple in the electron transport chain (ETC) whether uncoupling represents roles of mitochondria place them at the (Fig. 2). This membrane potential mitochondrial dysfunction (damage) or centre of the cellular mechanisms provides the driving force for protons a regulated cellular response to stress responsible for a growing number of to flow back into the matrix, which (UCP) in ageing muscle. pathological conditions. Despite an drives ATP synthesis by complex V One mechanism for increased explosion of research into (F1F0 ATP synthase). Protons may also mitochondrial function in ageing and bypass the ATP synthase and leak mitochondrial proton leak with age is diseas,e there are important issues that across the inner mitochondrial that oxidative damage to mitochondrial remain unresolved. One of these is the membrane (IMM), short-circuiting the proteins and lipids leads to membrane damage making the IMM more significance of the coupling efficiency coupling of ATP synthesis and O2 of oxidative phosphorylation (P/O). consumption. This leak will dissipate permeable to protons. The the membrane potential and reduce the accumulation of oxidative damage to Reduced mitochondrial coupling in efficiency of oxidative phosphorylation mitochondria is well documented in ageing tissues presents an interesting (lower P/O). aging tissues and damage to membrane paradox. Reduced coupling has been lipids has been shown to increase demonstrated to reduce the generation Increased proton leak is the most well proton leak in isolated systems. of ROS by the mitochondria. However, accepted mechanism to explain the Further support for a role for oxidative uncoupling with age is also associated lower P/O values in aged muscle. This damage in mitochondrial uncoupling with impaired ATP synthesis and hypothesis is supported by results comes from transgenic mice with altered cell energetics. Therefore, demonstrating increased proton leak in reduced antioxidant activities. certain changes associated with reduced coupling, such as reduced ATP levels, may be detrimental to cell survival, while others, like a decrease in ROS production, may be an adaptive response to cellular stress. Here I focus on mechanisms and significance of reductions in mitochondrial coupling efficiency in ageing muscle.

We have recently demonstrated significant mitochondrial uncoupling in vivo in aged mouse skeletal muscle (Marcinek et al. 2005). Using a combination of magnetic resonance and optical spectroscopy to directly Figure 1. Severe uncoupling in aged mouse skeletal muscle. Mitochondrial O2 consumption and ATP synthesis measure ATP use and O2 uptake in vivo, were determined in vivo in resting mouse hindlimb muscle. The ratio of ATP produced per O2 consumed (P/O) we found that mitochondria in aged was approximately 50% lower in the old (30-month) mouse muscle than in the young (7-month) mice (* P < 0.05). mouse skeletal muscle produce on Data from Marcinek et al. (2005).

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Mitochondria isolated from these mice had higher levels of oxidative damage and lower respiratory control ratios (state 3/state 4 respiration) (Williams et al. 1998), which is indicative of increased proton flux across the IMM.

In contrast, an increase in mitochondrial proton leak with age may be due to an upregulation of UCP activity. Increased proton leak through the UCPs would lower the membrane potential and lead to reduced ROS Figure 2. Schematic model of oxidative phosphorylation showing paths for uncoupling of oxidative production by the ETC, because phosphorylation. Oxygen consumption by the electron transport chain (ETC) and ATP synthesis in the mitochondrial ROS production is very mitochondria are coupled by the membrane potential created by ETC proton pumping across the inner sensitive to changes in the resting mitochondrial membrane (IMM). The ETC also produces reactive oxygen species (ROS) that cause oxidative stress and can lead to uncoupling. In the case of mild uncoupling, oxidative stress leads to upregulation of proton membrane potential. This means that leak through the uncoupling proteins (UCP). The increased leak reduces ROS generation and decreases the even a small amount of proton leak due oxidative stress. When oxidative stress exceeds cellular defenses oxidative damage can accumulate causing to UCP activity could lead to large membrane damage and severe uncoupling. decreases in ROS production. indirect evidence for a protective effect levels may then push the cell toward This idea has led to the proposal that of mild uncoupling. apoptotic cell death and lead to tissue mitochondrial uncoupling is actively degeneration. Under this scenario, regulated through UCP function as a In contrast, the more severe uncoupling differences in UCP expression and the defence against oxidative damage to that has been found in aged mouse degree of mild uncoupling would mitochondria. Uncoupling by UCP3 in (Marcinek et al. 2005) and human indicate differences in defense against muscle has been found to increase in (Conley et al. 2000) muscles in vivo is oxidative damage and contribute to the response to lipid peroxides (Echtay et more likely associated with variation in mitochondrial dysfunction al. 2003). Since the lipid peroxides mitochondrial damage and dysfunction. and tissue degeneration found in form in the mitochondria in the The presence of mitochondrial different tissues. presence of increased oxidative stress dysfunction is indicated by reductions in the cell’s energy state (ATP/ADP) Acknowledgements their role in upregulating UCP3 activity Thanks to Kevin Conley and Martin and/or ATP levels that accompany the supports a feedback model where Kushmerick for helpful comments lower P/O values in both species. The oxidative stress leads to mild during the preparation of this change in the cellular energetics is a uncoupling, which in turn reduces ROS manuscript. Development of these sign that the ability of the mitochondria production. ideas was supported by NIH grants to meet the energetic demand of the cell AG00057, AG022385 and AR36281. The oxidative damage and regulation of is compromised in these tissues. UCP hypotheses are not mutually Because reduced ATP levels can be David J Marcinek exclusive. This raises the interesting signal for apoptosis, the severe Department of Radiology, University of Washington possibility of different degrees of uncoupling may be an early step in the Medical Center, Seattle, WA, USA pathway toward tissue degeneration. uncoupling in aging tissues – one that References may be a regulated cellular defence Conley KE, Jubrias SA & Esselman PC (2000). Oxidative capacity against oxidative damage, and one that In the model presented here UCP­ and ageing in human muscle. J Physiol 526, 203-210. mediated mild uncoupling acts as a is the result of mitochondrial damage. Echtay KS, Esteves TC, Pakay JL, Jekabsons MB, Lambert AJ, In the first case mild uncoupling would regulated, adaptive response to reduce Portero-Otin M, Pamplona R, Vidal-Puig AJ, Wang S, Roebuck SJ & mitochondrial oxidative stress to Brand MD (2003). A signaling role for 4-hydroxy-2-nonenal in be due to upregulation of UCP activity regulation of mitochondrial uncoupling. Embo J 22, 4103-4110. in response to oxidative stress. This minimize oxidative damage. When mitochondrial oxidative stress increases Harper ME, Bevilacqua L, Hagopian K, Weindruch R & Ramsey JJ would create a negative feedback loop (2004). Ageing, oxidative stress, and mitochondrial uncoupling. Acta by lowering the membrane potential with age or in disease states, eventually Physiol Scand 182, 321-331. the level of oxidative stress exceeds the and reducing ROS production. Marcinek DJ, Schenkman KA, Ciesielski WA, Lee D & Conley KE Speakman et al. (2004) found that ability of the cell to counteract this (2005). Reduced mitochondrial coupling in vivo alters cellular proton leak was greater in skeletal stress and the result is damage to energetics in aged mouse skeletal muscle. J Physiol 569, 467-473. muscle mitochondria isolated from the mitochondrial DNA, proteins, and Speakman JR, Talbot DA, Selman C, Snart S, McLaren JS, Redman lipids. This damage accumulates P, Krol E, Jackson DM, Johnson MS & Brand MD (2004). longest-lived mice in a population. Uncoupled and surviving: individual mice with high metabolism have They demonstrated that UCP3­ leading to mitochondrial dysfunction, greater mitochondrial uncoupling and live longer. Aging Cell 3, 87-95. including the severe uncoupling we mediated uncoupling accounted for a Williams MD, Van Remmen H, Conrad CC, Huang TT, Epstein CJ & significant fraction of the difference have identified in vivo in aging skeletal Richardson A (1998). Increased oxidative damage is correlated to muscle. The associated disruption of altered mitochondrial function in heterozygous manganese between the longer- and shorter-lived superoxide dismutase knockout mice. J Biol Chem 273, 28510­ individuals. This study provides cellular energetics and reduced ATP 28515.

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Granny’s going global… Investigating the organisational and functional principles operating within the social brain involves establishing causative links between behaviour and multi-component neural systems. With the introduction of new techniques for monitoring brain activity, are we now advancing towards an understanding of how brain systems mediate social cognition? Alister Nicol and colleagues look at current strategies and some of the challenges lying ahead

So how do our brains achieve this? And individually recorded neurons. One how do we find out? major finding is that brains may encode complex visual stimuli, such as In humans, non-invasive techniques for complete faces (e.g. Young & Yamane, monitoring brain activity such as fMRI, 1992; Sugase et al. 1999) or an MEG and EEG, in the context of individual’s specific odour (e.g. individuals carrying out visual or Kendrick et al. 1992), using very few olfactory recognition tasks, have neurons – ‘Grandmother neurons’, revealed the topography and global named after a hypothetical cell organisational aspects of the underlying encoding Granny. However, brain systems. However, interactions grandmother neuron schemes (also amongst individual neurons within the known as ‘sparse codes’ or ‘local Alister Nicol (left), Hanno Fischer, Andrew Tate and neuronal networks involved have yet to encoding’) are not the only strategy Keith Kendrick. be revealed. brains use to encode complex stimuli: for example, odours (often mixtures Progressing in our career, finding a Our current understanding of encoding with hundreds of components) are partner, leading a social life – living in strategies within such networks has globally represented across a cell a modern society is a complicated been determined from the responses of population, each cell responding in its business. It is all about ‘how to play the game’, which ‘pack to join’, whom to interact with and how, in order to succeed. We are well equipped for this task: our brains possess specialised neural systems for perception, processing, assessing and memorising the vast amount of sensory information related to individual identity, emotional state and attraction, the interpretation of which guides our social behaviour. These neural systems are not unique to us humans and have been demonstrated in many social mammals including monkeys, ungulates, and rodents (Tate et al. in press).

Many species use visual cues to identify one another and communicate social signals, with the face playing an important part in human and primate societies. Others, like rodents, use odour cues for individual recognition. While the sensory receptors in our eyes and noses are exquisitely sensitive in detecting and conveying visual and odour cues from other individuals to Figure 1. A, Multi-electrode array recordings in sheep inferior temporal cortex (red grid in insert). Simultaneous our brain, it is the brain which must recordings were made from 238 neurons during a face recognition task. Here two presentations are made of a decode the constant stream of picture of the face of a sheep (bottom trace). Circles indicate a ‘hot-spot’ of activity. Spike frequencies of individual information, interpret its relevant neurons (in spike sec-1, [Hz]) are represented in pseudo-colour. B, Presentation of an odour evokes a specific features, and memorise and recall them pattern of activation across an array in the rat olfactory bulb. Each cell in the grid represents a change in the activity of a single neuron at a given location on odour presentation. Comparison of the array response in ii to that in a wide range of different social in i reveals that response specificity for a single odour may be supported by only a sparse sub-set of neurons contexts. (towards the red end of the spectrum in iii).

Physiology News | No. 63 | Summer 2006 | www.physoc.org FEATURES PN 33 own way with greater or lesser reproducibility (Laurent, 2002). The neural code for Granny’s face or smell, say Chanel No 5, is thereby held in the cross-cell activity patterns of the neuronal population (known as population or global encoding). Theoretically, the number of stimuli that can be encoded by an ensemble of neurons using population encoding increases exponentially with ensemble size, and so is much greater than using grandmother cells, in which the number of stimuli encoded increases only linearly with population size (Rolls et al. 1997).

However, the process of perception is Figure 2. A, Changes in array responses of neurons in the sheep inferior temporal cortex related to socially only one side of the problem. Our relevant stimuli such as different faces (see i). (ii) In the example shown, subtraction of activity maps recorded current knowledge on how brains during the presentation of two different face results in an array difference of 8%. This suggests that a successful discrimination between two faces can be achieved with very small changes in the networks. This also implies that ‘streamline’ the influx of sensory they are capable of representing many different individual faces. Data represent normalised spike frequency information and extract socially changes (adapted from Tate et al., in press). B, Neurons across an array may be incorporated in various ways important features such as identity and into a response. Here, spikes were sampled from 4 neurons (A–D) at four different electrodes in an array in the rat emotional state of individuals post­ olfactory bulb. In pre-stimulus activity, neurons A, B and D are synchronised – they are prone to firing at approximately the same time, for example in the grey band. During odour presentation, neuron B becomes perception is still inconsistent (Calder desynchronised from A and D. Relative firing rates pre- and during stimulus presentation (right) show neurons B & Young, 2005). Are high-capacity and C increasing their firing rates in response to odour presentation, while neuron A’s activity decreases. In this encoding schemes involving entire respect, neuron C does not respond. A recurring sequence (DABC) incorporating spikes from all four neurons networks of neurons used during the occurs more frequently during odour presentation than in pre-stimulus activity. So neuron C, whilst neither ‘responsive’ nor synchronised with any of the other neurons, is nonetheless involved in stimulus encoding. initial process of streamlining, whereas grandmother schemes represent a low capacity but highly concise top-level techniques allow simultaneous ensemble of neurons and the amount of code for memorising and recalling? sampling from several hundred neurons overlap between individual across a relatively wide area of brain representations; (2) activity ‘hot-spots’, The biggest obstacle in answering these tissue (several mm2). Data capture rates i.e. areas of neurons in close proximity questions is that most brains consist of are large; in our laboratory, as much as changing their activity in response to billions of cells (e.g. an estimated 50 100Mb min-1. If implanted chronically, stimulus presentation (Fig. 1), and billion in humans) – too many to be recordings can be made over a period alterations in the ‘temperature’ and explored one by one. The output of any of several months. location of these areas through learning neuronal network is defined not only by and memory; (3) correlation analyses to the properties of its cells but even more In general, the large volume of data explore whether global stimulus by the interactions between those cells. requires that most labs develop data properties, such as identity or Addressing network function, therefore, processing techniques including expression, are reflected by the degree requires simultaneously monitoring the algorithms for discriminating the of synchronization between neurons; activities of many cells. However, most activities of individual neurons (each (4) stimulus encoding in spatiotemporal of the responsive neurons identified so channel of MEA data may contain the sequences of neuronal activity, i.e. far are scattered across rather large activity of several neurons) and ‘patterns-within-patterns’ (Fig. 2). areas of the brain. methods for statistical analyses (e.g. Horton et al. 2005) in order to What do we gain? Over the last decade, many laboratories determine the spatial and temporal (ours included) have invested relationships between them, and so Many current hypotheses regarding considerable time in developing decipher the neuronal code. network function underlying sensory techniques to monitor the activity of encoding can only be tested by large ensembles of individual neurons, Our current research focuses on (1) the recording the activities of many e.g. using real-time calcium imaging in contribution of individual cells in an neurons simultaneously. Furthermore, transgenic mice. Another more widely ensemble to the overall ensemble computer simulations (invaluable in used approach is multi-electrode array pattern of response to defined visual understanding complex systems such as (MEA) electrophysiology (e.g. stimuli such as faces, facial expressions brains), inevitably rely upon sufficient Nicolelis & Ribero, 2002). Depending or odours. This includes overall array supporting biological data that are on the array size (in our lab we use differences to analyse the reliability of arrays of up to 128 electrodes), MEA a representation across a recorded (this article concludes on p. 50)

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‘Specific criticism' out whether ‘Heart throbs are real neuronal control mechanisms have The popular press often falls into the sparks or mere transients?’ could be of evolved independently from each other. trap of garbling the convention for interest to the addicts of reality TV. species names i.e. Forename-surname There could also be sting operations (so Our session brought together leading (upper case, lower case) plus italic popular in this country) depicting researchers who dealt with topics that letters. The most common offences are physiology, philately and philandering spanned across model systems (insects, again E(scherichia) Coli , in the recent meeting of The Society or molluscs, primates, carnivores) and Tyrannosaurus Rex and Homo Sapiens. secretly recorded rantings and methods (neurophysiology, behavioural But now Physiology News has managed chantings of drunken scientists in the physiology, biomechanics and modelling). it in the item reporting the Nobel Prize wine and cheese party. Philosophical One recurring theme was that animals award to Robin Warren and Barry discourses, like’ When does the life utilise many ‘tricks’ to simplify the control Marshall (62, 39). So, that should be begin?’ by the President of newly of movements. For example, Binyamin Helicobacter pylori and not formed ‘Society for Prevention of (Benny) Hochner described how, during Helicobacter Pylori [sic]. Since it Cruelty to Stem cells’ is certain to object retrieval, the highly flexible arm of becomes H Pylori through the rest of attract mature insomniacs desperately an octopus forms almost static bends that the item, PN can perhaps claim seeking an afternoon nap. Quiz contests act rather like joints, and thus reduce the consistency if not accuracy. between teams of smart, sweet and effective degrees of freedom of the savvy purveyors of physiology, movement. Tom Matheson described how Another detail from Physiology News representing lifers in Liverpool or 62, 3 – the explanation of the cover Manchester Un-united are sure to high levels of joint stiffness can facilitate photos [sic] implies we could have enthrall the audience. Yes Mark, you load compensation in locusts, and Volker expected four items of Brazilian have unleashed a Cain mutiny! Dürr demonstrated that aspects of wildlife, rather than just the one stepping, searching and reaching glorious iguana (and that'll be the J Prakasa Rao movements in an insect can in principle Green iguana, Iguana iguana, I guess) Department of Physiology, Christian Medical College, be generated by a single control network. that adorned the actual cover. Wot, no Vellore, India. In other invited talks, Paul Gribble Mardi Gras images from Rio? But I described how humans represent joint eventually discovered the remaining interaction torques, Trevor Drew provided three on the back page; very beautiful Goal-directed limb evidence that the neural mechanisms they are too. That's capybara (sing.) and movements underlying reaching in cats may have capybari (pl.) for Hydrochoerus developed from those used to control hydrochaeris, by the way. And is The Society for Experimental Biology aspects of locomotion, and Tamar Flash Main Meeting was held at the University 'caimen' the plural for the caiman provided a wide-ranging talk on of Kent at Canterbury from 3–7 April. A (Caiman latirostris)? computational issues in motor control. symposium on goal-directed limb David J Miller movements was organised by Tom The talks were well attended, and Institute of Biomedical and Life Sciences, University of Matheson (University of Leicester, UK) feedback was uniformly very positive. A Glasgow, UK and Volker Dürr (University of Bielefeld, general discussion provided a valuable Germany). forum for the development of some Cain mutiny? Goal-directed limb movements are interesting ideas, some of which were When I showed Mark Cain’s article intrinsic to many behaviours, in animals mulled over at more length during a very (Physiology News 62, 40) to my friend ranging from insects to humans. Making enjoyable evening meal attended by all of Batha Khani, host of the famous Indian such movements requires the solution of the contributors. A number of posters TV show Spat Chat, he made several problems as diverse as coping with associated with our symposium generated interesting suggestions to ensure high redundant degrees of freedom, planning considerable discussion during the main viewer ratings for physiologists. Juicy and controlling hand or foot trajectories, poster session. titles are necessary he felt, to draw the integrating biomechanical properties of attention of people. A series called Generous funding provided by the the limb, and compensating for postural Escape Artists could feature Company of Biologists (through the SEB), changes that affect balance. An important physiologists with distaste for The Physiological Society, and the goal of current neuroscience research is temperate environs, to star in episodes German Zoological Society (DZG) to develop an understanding of the like Cute Coote on high cliffs, Musings enabled us to support all of our invited sensory-motor transformations that during freezing: little igloo on the speakers, to award travel grants to two underlie these movements. A comparative glacier or Devilish dive of Neil’s students who presented posters and to approach can reveal general principles of Donald Duck. Debates showing the three selected presenters of Dubious Raymond pitted against limb movement control, because animals submitted talks. Timeless Hawkins discussing the as different as an octopus, a locust or a metaphysics of muscle or Denis the cat must all solve the same problems but Tom Matheson Menace facing Sandy Kenters to find their limbs and the corresponding University of Leicester, UK

Physiology News | No. 63 | Summer 2006 | www.physoc.org REPORTS PN 35

research concerning the role of nitric ‘Northern’ Cardiovascular oxide and the various NOS isoforms in Research Group regulating myocardial function. A topical and intriguing subject with a The 14th annual meeting of the plethora of interactions for those ‘Northern’ Cardiovascular Research interested in the regulation of Group was hosted by CRISTAL (the excitation-contraction coupling in the Cardiovascular Research Institute at heart. Leeds) in the Worsley Building at the University of Leeds (pictured right) on The day’s proceedings concluded with 12 January. The meeting serves as a a poster session which gave everyone a forum for the exchange of current chance to present and discuss their research between those working in the latest work. There was a wide diversity cardiovascular field, and this year we been identified, but areas of of research on show, and the posters saw researchers gathering from all over considerable interest are still being provoked such intensive discussion the UK – Manchester, Liverpool, investigated and the study certainly around the room that no one seemed to Oxford, Bristol, and Glasgow. showed once again that the familiar notice when the food started to be spectres of smoking and obesity reign served. A series of shorter talks by group supreme in shaping the risk of heart members was peppered by more in­ disease. After lunch, our guest It was great to have such a diversity of depth presentations. It was a privilege international speaker Ernst Niggli excellence in cardiovascular research in to have the British Heart Foundation’s (Bern) gave a comprehensive overview one location for the day. Thanks are Professor Stephen Ball (Leeds) present of his pioneering work on the control of due to the British Heart Foundation, the highlights of one of the most calcium release from the sarcoplasmic The Physiological Society and Leica ambitious cardiovascular studies reticulum in the cells of the cardiac for their generous sponsorship which undertaken yet in the UK. The British ventricle. During a sparkling made this meeting possible. Heart Foundation Family Heart Study presentation, with the odd wave thrown was first proposed in 1996, and it was in too, the latest ideas on how Matthew Lancaster fascinating to see how it had developed intracellular calcium control is School of Sport and Exercise Science in its attempt to identify genes that regulated were delivered. Concluding Sarah Calaghan directly correlate with the risk of heart the talks for the day, Barbara Casadei Institute of Membrane and Systems Biology disease. No definitive genes have yet (Oxford) gave insight into her extensive University of Leeds, Leeds, UK

Physiology News | No. 63 | Summer 2006 | www.physoc.org 36 PN REPORTS

NC3Rs Stakeholder Meeting

The National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) held its second Annual Stakeholder Meeting in London at the end of January. This was a chance for interested stakeholders to hear how the Centre had progressed over the previous 12 months and what plans it has for maintaining the momentum on raising the profile of the 3Rs.

The NC3Rs is an independent scientific organisation that was set up by the UK Government in 2004 to act as a catalyst for the 3Rs. It brings together Lord Turnberg, Chair of the NC3Rs Board academia, industry, Government, regulators, and animal welfare and protection groups with the aim of The Stakeholder meeting was a chance training. One project already completed advancing the 3Rs through funding for the NC3Rs staff, and the scientists is the production of a DVD on research, providing access to that they are working with, to report on refinement in primate husbandry by a information and fostering the growing list of activities that the group from Oxford University. Clips collaborations. Centre is now involved in. The NC3Rs from the DVD are now available to main funding scheme awarded a total download from the NC3Rs website. The NC3Rs has an extensive website, of £1million to eight projects. which includes an Information Portal to Successful applicants included one The Centre takes a scientific approach help scientists apply the 3Rs by developing a cell-based diabetic wound to looking at where numbers of animals providing access to the most useful and bioassay and another looking at can be reduced and brings together relevant references. This sits alongside reducing animal usage by multiple groups of scientists, both from details about all the Centre’s funding antigen immunisation schedules. academia and industry, to think about schemes and activities. An e-newsletter opportunities for doing this. is issued on a monthly basis to There is also a small award scheme, run highlight new content on the website jointly with LASA (Laboratory Animal One example is an initiative on and can be subscribed to through the Science Association), which gives up to experimental design which is being led site. £2k for small projects, visits or by Michael Festing. A study is being carried out, in collaboration with the National Institute of Health, to assess the quality of experimental design and analysis in a sample of 300 papers reporting research involving animals. It is hoped the study will be completed by the end of the year.

The meeting also saw the inaugural 3Rs Research Prize being awarded by Lord Sainsbury. The recipient was Siouxsie Wiles of Imperial College London for her work on refining studies of gut infection in mice. She also received £10k to continue applying the 3Rs in her work.

Tim Watson Communications Manager, National Centre for the Replacement, Refinement and Reduction of Animals in Research, London, UK

Siouxsie Wiles receives the inaugural 3Rs Research Prize from Lord Sainsbury www.nc3rs.org.uk

Physiology News | No. 63 | Summer 2006 | www.physoc.org REPORTS PN 37

Moving forward with neuromuscular transmission

The neuromuscular junction (NMJ) provided the very first insights into synaptic transmission and continues to yield exciting new data, both as a model synaptic system and as the key to normal and dysfunctional motor control. This was evident at a one Symposium speakers (left to right) : Jack McMahan (Stanford), Peter Molenaar (Leiden University Medical day symposium held in Newcastle on Centre), Hans Brenner (Basel), John Harris (Newcastle), Clarke Slater (Newcastle), Terje Lømo (Oslo), Guy the 28 October 2005 to mark the Bewick (Aberdeen), Hugh Willison (Glasgow), Angela Vincent (Oxford) and Richard Ribchester (Edinburgh). retirement of Clarke Slater, jointly sponsored by The Physiological neuromuscular synaptic protection in that by using variable tilt angles and Society and the Institute of the Wlds mouse mutant in which 3D reconstruction it was possible to Neuroscience at Newcastle. Clarke Wallerian degeneration is perform high resolution electron has dedicated most of his research dramatically delayed. This has microscopic tomographic studies of career to the study of the NMJ and triggered a search for other genes the frog NMJ with a spatial resolution the speakers who contributed to this which confer neuroprotection through of 2-3 nm. This has revealed the symposium included many of his the expression of mutant Wlds protein stereotypic orientation of docking long standing collaborators. Under in the motor neurone. proteins on the synaptic vesicle the title Moving forward with membrane which are associated with neuromuscular transmission, this was With respect to motor neuropathies, a highly organised arrangement of not so much a reflective day as a such as myasthenia gravis (MG), it is membrane-bound filamentous review of the current state of the field clear that many of these arise from macromolecules found on the pre­ and a perspective of the future autoantibodies directed against ion synaptic membrane (active zone challenges based around two major channels, receptors or associated material). It has been possible to themes of synapse formation, membrane proteins. In this respect visualise the rotation of undocked plasticity and degeneration at the Peter Molenaar (Leiden) and Angela vesicles in order that they become NMJ (Bewick, Brenner, Lømo and Vincent (Oxford) described how correctly orientated to the orderly Ribchester) and pathologies and antibodies from patient groups could array of macromolecules on the toxins which affect the NMJ (Harris, affect NMJ function through targeting synapse. This arrangement helps Molenaar, Vincent and Willison). the acetylcholine receptor (AChR) or maintain the vesicles and calcium MuSK, a tyrosine kinase receptor channels at precise distances and, Both formation and degeneration of essential for the agrin-induced thereby, plays a major role in the synapses are known to involve rapsyn-dependent clustering of vesicle fusion process. extensive reciprocal interactions AChR. In some instances, inhibiting between motor neurone and muscle AChR function by binding with low Despite the great advances that have fibre to direct synaptogenesis and affinity acts allosterically to increase taken place during Clarke’s career it plasticity. The central role of agrins desensitisation. In contrast to AChR­ is clear that the NMJ has not yet in this process is well established, but related neuropathies, Hugh Willison given up all its secrets and will Hans Brenner (Basel) showed recent (Glasgow) described how murine continue, in the late Sir Bernard data that neuroregulin is dispensable models have revealed the role of Katz’s words to be ‘an experimentally and that, in some instances, the early GQ1b ganglioside antibodies to favourable object whose study could stage formation of the post-synaptic induce complement-mediated NMJ throw considerable light on synaptic specialisation can occur without degeneration. Such delineation of the mechanisms elsewhere’. agrin, challenging the ‘neurocentric’ antibody-dependent processes will view of NMJ formation and have clear benefits for design of Colin Ingram suggesting some autonomous therapeutic interventions. Director, Institute of Neuroscience, Newcastle University, UK organisation within the muscle fibre. The complex processes that maintain Finally, the enormous insights that Richard Ribchester the NMJ were illustrated by Richard can come from studies of functional Centre for Neuroscience Research, University of Ribchester (Edinburgh) who anatomy were illustrated by Jack Edinburgh, UK described neurodegeneration and McMahan (Stanford). He showed

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matched with the functional demands of tissue, mainly in skeletal muscle. The second involves cardiovascular control, largely concerned with the Introducing Editors dysfunction associated with hypothermia (see Physiology News 60, appointed in 2006 12).

Margaret Brown He has served on the Council of the Maggie is a Reader in Cardiovascular British Microcirculation Society and Physiology in the School of Sport and the Society for Experimental Biology, Exercise Sciences at the University of and recently retired as Convener of the Birmingham, where her current Comparative Physiology SIG. research interests lie in the mechanisms of vascular and microvascular Simon Langley-Evans remodelling in skeletal and cardiac Simon is Chair in Human Nutrition at muscle, from the molecular basis to the University of Nottingham, UK. His integrated physiology and therapeutic research is focused on the application. The role of exercise as a developmental origins of health and modality for manipulation of blood disease. Using animal models in which vessel function and growth is the maternal diet is altered during particularly pertinent in cardiovascular pregnancy, he is studying the long-term and ischaemic diseases, involving changes in renal and vascular studies of patient cohorts undertaking physiology of the resulting offspring training programmes. and the molecular basis of developmental programming. Jim Deuchars Jim is Professor of Systems Paolo Maddedu Neuroscience at the University of Paolo is cardiologist and chair of Leeds. His current research interests Experimental Cardiovascular Medicine focus on the properties of neurons and at the Bristol Heart Institute, University circuits participating in control of of Bristol. His research, funded by autonomic functions. charities and the European Community, focuses on therapeutic angiogenesis Stuart Egginton using growth factors and stem cells for Stuart is a Reader in Physiology in the the cure of ischaemic complications Centre for Cardiovascular Sciences at with special emphasis on diabetes. The the University of Birmingham. Born in group leaded by Prof. Madeddu has the North of England, he has discovered the angiogenic property of appreciated the opportunity for travel human tissue kallikrein, nerve growth associated with a career in science: a factor, and PAR2. BSc in zoology from the University of Wales (Bangor), then a PhD in Simon Malpas physiology from the University of St Simon is an Associate Professor in the Andrews in Scotland (working with Ian Department of Physiology and the Johnston), followed by a personal Bioengineering Institute at the Fellowship for post-doctoral studies at University of Auckland, New Zealand. the University of Maine in the USA (in His primary research interest lies in the lab of Bruce Sidell), before understanding the role of the returning to the UK for a post-doc at sympathetic nervous system in the Birmingham (with Olga Hudlicka), development of hypertension. His where he has remained. Appointments New members of the research interests include neural control in Austria and New Zealand have of the circulation, medical device further broadened his scientific and Experimental Physiology development and telemetry. geographical horizons. Editorial Board, from the top: Margaret Brown, Jim Kaushik Patel Stuart has two main areas of research Kaushik is a Professor in the Deuchars, Stuart Egginton, interest, with a degree of overlap that Department of Cellular and Integrative has varied with time. The first is how Simon Langley-Evans and Physiology at the University of remodelling of the microcirculation is Paolo Madeddu. Nebraska Medical Center in Omaha.

Physiology News | No. 63 | Summer 2006 | www.physoc.org THE SOCIETY’S JOURNALS PN 39

His general research interest is in Javier Stern neural control of the circulation in Javier is an Associate Professor in the health and diseases such as heart Department of Psychiatry at the failure, diabetes and hypertension. His University of Cincinnati, USA. His current focus is on the role of the general research interestx relate to paraventricular nucleus of the understanding the cellular mechanisms hypothalamus in the control of fluid controlling neuronal excitability, balance. synaptic connectivity and plasticity in central neuronal circuits involved in Michael Shattock autonomic and neuroendocrine control, Michael is Professor of Cellular including ion channel modulation of Cardiology at King’s College London. electrical activity of hypothalamic He runs The Cardiac Physiology neurones, neuronal structural and Research Group within the functional plasticity in health (exercise, Cardiovascular Division of King’s lactation) and disease (hypertension, College London. The Group’s general heart failure, diabetes) conditions. interests lie in studies of the regulation and function of ion translocating Michael White proteins in the cardiac myocyte in As an undergraduate Mike read health and disease. The group exploits physiology at the University of Leeds a range of models and techniques and then worked in the MRC including molecular and protein based Environmental Physiology Unit in studies, single cell electrophysiology, London before completing a PhD at the cell cultures, isolated muscles and University of Nottingham. In 1986 he isolated hearts. Current interests of the moved to Birmingham and the then group include (i) the regulation of Na/K newly formed School of Sport and ATPase by the accessory protein Exercise Sciences, where he has phospholemman, (ii) the role of T-type remained ever since. Now Reader in Ca channels in cell growth and Exercise Physiology he has worked on proliferation, (iii) signalling in ischemic human thermoregulatory and exercise preconditioning in adults and neonates physiology, the effects of disuse and and (iv) regulation of signalling and ion ageing on human muscle function and translocating by thiol modification and human cardiovascular control during oxidant stress. exercise in health and disease.

New members of the Experimental Physiology Editorial Board, from the top: The Editorial Board of Experimental Physiology now holds one of its two meetings each year in the USA to coincide with the FASEB Experimental Biology Meeting and to Simon Malpas, Kaushik facilitate attendance by the increasing number of overseas Editors. Patel, Michael Shattock, A good percentage of the Editorial Board, including five new Editors, were able to attend Javier Stern and Michael the meeting held this year on 3 April in San Francisco (photo by Prem Kumar). White.

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A century (and more) of physiology now online

Early in May the complete scanned archive of The Journal of Physiology finally went live on PubMed Central (PMC).* The scanning and checking of around 15,000 articles (no-one has actually counted them) published between 1878 and 1997 has taken longer Then and now – The Journal of Physiology in 1878 than expected, but the results are and 2006 (above) and the development of impressive. Within a few months the Experimental Physiology over the years (left). remainder of the archive, from 1998 to Journal of Experimental Physiology 2005, will also appear on PMC. At that (1908–1936) I would like to hear of it so point the only content not present on the that we can update the contents lists.) PMC site will be the most recent year’s issues which are available to subscribers Experimental Physiology contains its on HighWire and Synergy. issue of Physiology News by John Coote share of classic papers. John Coote’s and Thelma Lovick (62, 16). Top Ten papers list begins with (1) The Wellcome Trust has provided most Sherrington C S (1908). Some of the funding for the archive project, The scanned archive is by title, author, comparisons between reflex inhibition part of a joint initiative with the Joint institution, references and abstract and reflex excitation. Q J Exp Physiol 1, Information Systems Committee to keywords, and a PDF of the full paper 67–78; (2) Barrington F J F (1925). The digitise the archives of historically can be downloaded. The front and back effect of lesions of the hind-brain and important medical journals. A launch matter – editorial board lists, indexes, mid-brain on micturition in the cat. Q J party to celebrate the publication of the notices and advertisements – are Exp Physiol 15, 81–102. (3) Liddell E G archive was held on 11 May at the available through links at the foot of T (1953) (Sharpey-Schafer lecture). The Wellcome headquarters in London each issue’s contents list and are brain and muscle management. Q J Exp (pictured on p. 51), attended by various interesting to sample. In 1934 CF Physiol 38, 125–137. Officers of The Society and staff from Palmer (London) Ltd, Makers of The Society’s Publications Office in Physiological and other Scientific But there is also plenty to interest the Cambridge. Apparatus, were selling their Rubber more generalist reader within the lists Bellows Respiration Unit for £14 15s, for each year. In a paper on The action Aside from the great classic papers quite a price in today’s money. of tobacco smoke, with special reference familiar to every student, there are many to arterial pressure and degeneration by curiosities from the past to discover in A little earlier in the year the complete W Emerson Lee (1, 335–358), it is the archive. Try 54, 244–247 or 57, scanned archive of Experimental noted, more than half a century before 175–180 for a view of the questions and Physiology, formerly The Quarterly the dangers of smoking were taken technological difficulties that troubled Journal of Experimental Physiology, seriously, that ‘arterial disease may physiologists early last century. While was published on the HighWire site.† result from prolonged tobacco smoking’. many of the titles do not draw the casual This project was funded by The Society A review by McCance & Widdowson on reader in, some are intriguing. An article and organised by HighWire Press. The chemical structure of the body on The phosphurus distribution of published in 1956 (41, 1–17), based on resting fly muscle (80, 231–237) had me Like the J Physiol archive on PubMed the Scharpey-Schafer Memorial Lecture, wondering how many flies were Central, the Experimental Physiology provides, as well as a detailed account required for the experiments (‘enough’) archive is free to all readers and consists of nutritional studies up to that point, an and how they were caught (‘easily’; the of searchable PDF files of all papers insight into the prejudices and flies were reared in a cage which was back to 1908. There are still some perceptions of the time. then darkened while a beaker glitches to iron out on the site. The print Both archives offer a vast resource for illuminated at the back end was held at archive we obtained for scanning did not physiologists and all students of life the cage opening). Observations on the include information about months of science. Every week I get several composition of alveolar air on Everest, publication, and hence the repeated requests for copies of articles from the 1933 by Raymond Greene (82, months in many of the early years on journals that up until now have only 481–485) is a fascinating account of the archive page. (If anyone knows been available in departmental libraries. physiology in the extremes, to be where there is a collection containing It will be a pleasure to be able to refer contrasted with the report in the last the first 25 issues of the Quarterly readers to the archive sites. * http://www.pubmedcentral.gov/tocrender.fcgi?journal=236&action=archive † http://ep.physoc.org/contents-by-date Carol Huxley

Physiology News | No. 63 | Summer 2006 | www.physoc.org AFFILIATE NEWS PN 41

Babies and the bench In the last issue, Patricia de Winter questioned why there are so few senior women scientists. In this issue Dola Akanmu, a mother–of–two who has just obtained her PhD, examines the challenges that face women who have children and wish to continue in research

Bench work and babies is easy. In fact, parents – which can be helpful if you any work and babies is easy providing fall within the required income bracket. suitable childcare is in place and you This can reduce childcare fees by up to have a great boss who understands the 70% for the lower income bracket. In practical problems of having to literally reality, a significant reduction is only drop everything if your child becomes achieved providing your joint income is ill. This statement is, of course, well below £40k. dependent on many other factors: the type of experiments you do and When your child reaches the age of 3, whether your work schedule is flexible, up to age 5, they will be entitled to a for example. Some women have nursery voucher which is undertaken flexible working approximately £400 per term. This is arrangements that suit all parties most welcome particularly as the concerned, such as working from home paperwork is undertaken by the 1 or 2 days a week or doing 4 days on, childcare provider. The most recent 4 days off (as agreed by one employer). voucher scheme works through your Dola Akanmu – working in research whilst bringing up Mutually-agreed working arrangements two young children can be done – it just requires employer in combination with the allow you to be more focused, which is organisation, determination and a sympathetic Imagine Childcare Co-operative. particularly important as careful employer. Basically the employee can exchange planning of experiments is essential or ‘sacrifice’ their gross income for when time is at a premium. not favour the idea of travelling during childcare vouchers, and as this is rush hour, with a pushchair, baby bags, deducted from the gross salary, less Another factor to take into nappies and bottles in tow. income tax and national insurance is consideration is childcare, which can be payable. The maximum you can a big emotional change for both child The cost of childcare is an important sacrifice is £50 per week or £217 per and parent. If care provided by a consideration as a full-time place can month. Providing the scheme is open to relative is not feasible, then finding a be around £1,000 per month for the the employers of both parents, they can suitable au pair, childminder or nursery under 2s, depending on where you live. both claim, but there are no extra place may offer various benefits at In most cases this represents a large benefits if you have more than one different stages of the child’s life. Some proportion of one’s salary. To offset child in childcare. You can check employers have recognised that in­ these exorbitant childcare costs the whether your employers have signed up house nurseries offer particular benefits Government has provided some at www.childcarevouchers.coop. as the child is then on site should incentives to encourage women to anything untoward occur. However, if return to work. Family tax credits are Achieving a good work–life balance is you have to commute to work you may based on the combined salary of the a constant juggling act. Rushing away from work to collect your child means The Society’s journal archives that you do miss out on the social The Journal of Physiology archive from 1878 to 1997 is now freely available online aspects of work: drinks and seminars, at: http://www.pubmedcentral.gov/tocrender.fcgi?journal=236&action=archive which are very useful for generating ideas, networking and catching up with and Experimental Physiology archive from 1908 to 2004 at: any other business. Organising your http://ep.physoc.org/contents-by-date.0.shtml attendance at such an event involves See also A century (and more) of physiology now online (p. 40) and images from the Wellcome Trust launch careful advanced planning and hence party to celebrate the publication of The Journal of Physiology archive (p. 51) requires having adequate notice. Nevertheless working full- or part-time FRS for Experimental Physiology Consultant Editor in research whilst bringing up young Peter Hunter (right), Professor of Engineering Science and Director of the children can be done – it just requires Bioengineering Institute at the University of Auckland has been made a organisation, determination, and a Fellow of the Royal Society. sympathetic employer. Peter pioneered modelling the human body using specially developed algorithms plus methods that incorporate detailed anatomical and Dola Akanmu microstructural measurements and material properties into continuum King’s College London, UK models, most notably of the heart.

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his family – his wife, children and Henry Barcroft grandchildren – and he always spoke of To continue our occasional them with the greatest affection. He series of Memorable was lucky, especially in this day and Members, Ian Roddie praises age, that this affection was returned and they gave him their love and support the formidable intellect and right to the end. bonhomie of a much-loved Honorary Member Ian C Roddie See also Memorable Member – G L Henry Barcroft (1904–1998) was the Brown (Physiology News 62, 34). son of a famous father, Joseph Barcroft, one-time Professor of Physiology at Mistaken identity! Cambridge. Henry qualified at Cambridge with a double first in As many Members know, The Society physiology and worked at UCL with was founded in 1876 as a Dining Club. Lovatt Evans before moving to the In 1941, at a Society meeting in chairs of physiology in Belfast Cambridge, Charles Sherrington, one of (1935–1948) and St Thomas’s in understand it fully. It was when the the Founding Members, reminisced London (1948–1971). His best known young enthusiast was explaining his about those early days. Luckily a work was on nervous control of human proposition for the third or perhaps verbatim report is included in the short blood vessels and for this he was fourth time, with perhaps some account of The Society’s second 50 elected FRS in 1953 and an Honorary exasperation, that the flaws in the years produced in 1976 to mark The Member of The Society in 1974. proposition would become self-evident, Society’s Centenary (Bynum, 1976). as would the need for revision. Thus it Sherrington recalled that J S Burdon One of his most important and was possible for the proponent to Sanderson, another Founding Member endearing qualities was his ability to withdraw with dignity and self-esteem and first Waynefleet Professor in bring out aptitudes and enthusiasms in intact to think the proposition through Oxford, was notoriously absent minded. the people with whom he worked. Thus again. Henry would encourage this and He went on to say that he and eight or he catalysed the conversion of many say that he looked forward to hearing nine others were upstairs in the study at young graduates into professional the outcome. Burdon Sanderson’s home when physiologists. John McMichael, one Gaskell embarked on a long time Professor of Medicine at the Royal Henry’s courtesy and bonhomie made dissertation on his current theories. Postgraduate Medical School at him a much loved member of The After a time the dinner bell rang but Hammersmith, put it like this – ‘He is Society, but to some extent masked his Gaskell continued the argument; in the always surrounded by enthusiastic formidable intellect. The other­ end Lady Sanderson called upstairs that young men to whom he gives guidance worldliness of his outward demeanour dinner was served. Gaskell went and inspiration to a marked degree. tended to obscure a tough inner resolve downstairs followed by Sanderson and Completely unselfish himself, he gives to make the best of his opportunities in the rest. The dining room door was them every opportunity to develop their the real world. In making decisions, he open and Lady Sanderson already talents’. Sir Henry Dale, the famous could be quite ruthless and unswerving seated at the table. Gaskell, still Nobel Laureate, put it another way in his determination to do what he propounding his views, went to the when he said – ‘He has given abundant considered to be the right thing, head of the table, seated Sanderson on evidence of his power of awakening in regardless of the consequences. Once his right and began to carve; suddenly others an interest and enthusiasm his mind latched onto a problem, e.g. he looked up at his hostess, and said, comparable to his own’. the mechanism of functional ‘Pardon me, I thought I was in my own hyperaemia, he would pursue it house’. Sherrington added ‘That made Henry was a kind man – he could find relentlessly with a tenacity that was two absent-minded Members at the a good word to say about almost almost obsessional. He had an same time.’ anyone and character assassination was excellent memory for scientific, but totally foreign to his nature. (according to his wife) not domestic, Ann Silver detail. He stored in his mind a huge Nevertheless, he was a shrewd judge of Reference character and the strength of a scientific collection of snippets of scientific Bynum WF (1976). A Short History of the Physiological Society argument. When flawed propositions knowledge that would surface when 1926–1976. J Physiol 263, 23–72. were put to him by enthusiastic juniors, needed to interpret new data. he handled them gently and never Send us your recollections of past crushed or humiliated. He would listen Henry was fortunate in being born into Members of The Society for attentively but then ask for the a stable and loving family and this inclusion in our Memorable proposition to be explained again more good fortune was to stay with him Members series. Contributions to slowly so that his ‘aging brain’ could throughout his life. He was devoted to [email protected]

Physiology News | No. 63 | Summer 2006 | www.physoc.org PARLIAMENTARY AND SCIENTIFIC COMMITTEE PN 43

Parliamentary and Scientific Committee

The January meeting of the Parliamentary and Scientific Committee at Portcullis House, Westminster, covered the topic Hospitals of the future.

In his introduction, Sir Richard Sykes, Rector of Imperial College London, wirelessly connected hand-held devices. later discussion it was felt that selling postulated that hospitals of the future (Maybe we will even be able to keep this idea to the public was a major will revolutionise the way in which our mobiles switched on in hospitals). factor that would need to be carefully hospital care is delivered thanks to the argued, given the attachments of both impact of new technologies together Susan Grant (Projects Director, Atkins local populations and clinical staff to with major demographic and Design and Engineering Solutions, ‘their’ hospitals. Ms Grant indeed felt sociological change. He suggested that whom we thank for the illustrations of that eliminating the term ‘hospital’ new healthcare technologies, including future hospitals), an architect who would allow more constructive robotic surgery, wearable and specialises in hospitals, carried on this discussion, though this did have a touch implantable monitors exploiting theme, showing how hospitals in many of the Orwellian about it. wireless communication and new IT parts of the world were already systems will change both the incorporating new technologies in ways There was a frisson from the left­ environment and the staff base that were changing their architecture, leaning politicians present when Ara required. Diagnosis and treatment allowing creative approaches to Darzi mooted that patients’ perceptions would be more in the hands of building, particularly pre-fabrication tended to favour the private sector in specialised technologists rather than and the concept of the disposable or certain contexts. Apart from this there general physicians. Remote monitoring upgradable ward unit (the ubiquitous was remarkably little political would greatly change the location of term ‘pod’ was used both for room controversy, given the undoubtedly patient care towards a home base and units and mobile surgical theatres) to enormous costs of the new technologies hospitals would retain more specialist respond to changing technology and the involved and the consequences for the functions. challenges presented to hygiene by the individual of insurance companies threat of pandemics. knowing predictors of their health Richard Kitney, Professor of status, though data-security did emerge Bioengineering at Imperial College, Sir Ara Darzi of the Division of as a major concern. described the development of an Surgery at the, by now remarkably well For teachers of physiology, the England-wide medical database and represented, Imperial College, implications are interesting. The how the local intranet and internet discussed the impact of changing physicians and surgeons of the future together provided an integrated system technology on the culture of the will undoubtedly have to adapt to more for the storage and analysis of patient patient-professional interaction and specialised roles. General practitioners data and the potential for remote how alternative evaluations of clinical and others involved in community care diagnosis and even robot-based success by either prevention or cure will play a greater role in technology­ treatment. He commented that this had could have a major influence. He based diagnosis and treatment, as been greatly enhanced by the suggeted that the private sector would robotics and IT rolls this away from the development of generic components for deal with minor and elective treatments hospital. Their training will need to fit data storage and processing in many while the NHS would concentrate on them for this, perhaps by emphasising spheres of life, which was rapidly major treatments in specialist centres. the current trend towards learning how bringing down costs and putting He felt that the concept of the ‘local to learn rather than acquiring enormous power and access to data into general hospital’ had had its day. In the information. They might also need a harder edge to their scientific and technological knowledge. There will undoubtedly also be a need for high­ grade technologists, graduates with an undertanding of human biology and the flexibility to adapt to changing technologies. Our teaching of science and medicine will need to adapt.

Ian McGrath

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grant focuses on pancreatic acinar New Chief Executive physiology and pathophysiology, with a particular interest in the regulation of Michael Collis has been appointed as ion channels and Ca2+ signalling in The Society's new Chief Executive. He secretory cells. He has published will be responsible for helping to shape extensively in the field of physiology and deliver the strategic vision of The and has been a long-standing supporter Society, interacting closely with the of the discipline. Ole played key roles Trustees and Executive Committee and as a member of The Society’s Council maintaining well-established links with (formerly Committee) and as its other learned societies. We have been International Secretary from fortunate to have recruited an New Society President Ole Petersen 1992–1998. He will take up his post in established scientist with a long­ July 2006. standing interest in physiology and a Reviews European Editorial broad range of experience in academia Committee. Ole received his MB ChB Ole writes: ‘I look forward to working and industry. (1969) and MD (1972) from the with the Executive Committee and University of Copenhagen and was Council to further enhance the Michael has spent the majority of his appointed in 1975 as Symers Professor effectiveness and international research career in industry, where he of Physiology and Head of the reputation of The Physiological Society, held senior research management Department of Physiology at the which I have always regarded as my positions at Wyeth, ICI and Pfizer and University of Dundee. He was elected principal Society. advanced 16 novel drug candidates into as a Fellow of the Academy of Medical clinical development. As lead of the Sciences (FMedSci) in 1998, The Royal From my current work as European Academic Liaison and Collaboration Society in 2000, the Royal College of Editor of Physiological Reviews I know group for Pfizer, Michael established Physicians London (FRCP) in 2001 and that physiology can have a very high numerous links with academic has been the recipient of numerous impact and I therefore believe that the researchers and learned societies. He other prestigious awards and lectures. future is bright for our science as well has published extensively, particularly Ole Petersen's current MRC programme as our Society.’ on the role of purinergic receptors in the cardiovascular system and has served on the Editorial Boards of Alan North Hypertension and the British Journal of Council elected Alan North, FRS as Pharmacology. Michael is a strong President of The Physiological Society in believer in the importance of 2003 and he will be stepping down at our integrative physiology, and is currently AGM at University College London in July a member of the BBSRC Studentship 2006. I recall Alan's comments when he and Fellowships Committee, MRC took over as President from Colin Physiological Systems and Clinical Blakemore, FRS at the AGM in the University of Manchester - Alan stated that Sciences Board and the RAE (2008) Secretary David Eisner, was instrumental in he hoped to play a key role in the promotion Pre-clinical and Human Biological securing the International Union of of the discipline of physiology and The Sciences Panel. Physiological Sciences (IUPS) meeting for Society, especially in an era when traditional the UK in 2013. departments of physiology were now scarce. Michael Collis took up office in early Alan has certainly raised the profile of Moreover, he felt that The Society and its May and is based at The Society’s physiology and The Society. His wide range President needed to be pro-active in London Office. of physiological research interests and promoting the awareness and understanding extensive international experience, of physiology in academia, government, underpinned by the respect of scientific industry, and society at large, especially community for his research, have ensured New President young people who would become the that physiology remains at the forefront. physiologists of tomorrow. The Physiological Society is delighted Apart from his role as President of The On behalf of the Executive Committee, to announce the election of Ole Physiological Society, Alan is currently Vice Council and the membership, I would like to President and Dean of the Faculty of Life Petersen, FRS as The Society's new thank Alan for his invaluable advice during Sciences at the University of Manchester President. Ole is currently George Holt the past 3 years. As Chair of our Council and Chair of the Pre-clinical and Human Professor of Physiology and MRC meetings, Alan played a key role in Biological Sciences RAE 2008 panel. I have Professor at the University of Liverpool launching annual November breakout groups valued working closely with Alan and thank and holds honorary executive positions which not only reviewed The Society's him for his continued support and guidance as Vice-President of the Royal Society, ongoing activities but also identified strategy in the recent appointment of The Society's Secretary General of the International for the coming year. In addition to new Chief Executive, Michael Collis. Union of Physiological Sciences representing The Society in the UK and (IUPS) and Chair of the Physiological abroad, Alan together with our International Giovanni Mann

Physiology News | No. 63 | Summer 2006 | www.physoc.org BIOSCIENCES FEDERATION/SOCIETY NEWS PN 45

European Research Council (ERC) and that it inconceivable that scientists from 25 countries will have a budget of at least €1bn per annum. can have open access to the new ERC. The This will come from the FP 7 budget of €54bn organisation would sink without trace beneath for 7 years. You may be interested to know the many thousands of applications, and even that this budget is 40% higher than for FP 6. if these could be processed, with a high standard of peer review, the success rate I write that this is ‘good news’ because I think would be so low that the ERC would be it highly desirable that a UK bioscientist can discredited. have the opportunity to apply for response mode funding with a colleague from the The solution to this significant problem is mainland without having to go through the being sought and part of the solution may contortions that have been necessary in the come from reserving funds for ‘young Richard Dyer past. On this occasion, I think that the investigators’. There is currently a suggestion European Commission (EC) has been on the that 30% of the budget will be made available Much of the work of The Biosciences side of the angels! They have resisted all only to those just starting their careers. But Federation (BSF) is conducted through its attempts by national or special interest groups this will merely create another problem. standing committees. Currently these cover to dominate the ERC. As an example, the ‘Young’ means very different things in different education, animal science, environmental EC forbad the ERC Council from having 25 countries. If it is defined as 6 years ‘postdoc’ sustainability and Europe. One of their members! Any other number was allowed but then it means 30 in the UK but nearly 40 in responsibilities is to scan the horizon to look not one per state. There will be no juste quite a few other EC countries. for issues that may impact upon the retour: there was a time when ERC was biosciences because of new policies and going to mean European Research Whatever is decided, it seems inevitable to legislation. Often these changes will effect Competition. me that access to the ERC will have to be more than those of us with careers in biology restricted somehow. The BSF, through its – and this is especially true with developments All of this is very positive news – but there are European Liaison Group, will be closely on a European scale. some formidable problems ahead. First, think watching how this develops and we will about what science means in a European respond on your behalf when an appropriate In my view we failed to realise in advance the context. It covers 25 states and includes the occasion arises. full impact that the Fixed Term Directive would humanities and social sciences. Second, have on the employment of young people in think about the budget. €1bn is roughly the Richard Dyer our laboratories. You will know that today a same as the budget that our new Medical Chief Executive, Biosciences Federation first postdoctoral position can be regarded as Research Council can anticipate. Clearly, it is (http://www.bsf.ac.uk) ‘training’ but that a second 3 year postdoctoral position in the same laboratory usually makes the individual permanent. That is, he or she Bridget Lumb has tenure and will have to be made Bridget Lumb was elected as Meetings redundant at the end of the 6 years. As a Secretary of The Physiological Society in 2002 consequence, and even though the second and was the first female member of the ‘postdoc’ is often more productive than the Executive Committee under The Society's new first one, laboratory heads frequently now governance. Bridget has been extremely has now been adopted for our future Main undertake a cost benefit analysis when energetic in devising and implementing the new Meetings. thinking about keeping a ‘postdoc’ for a format for our Scientific Meetings. We now host second term. Whilst the cost of redundancy one Main Meeting and 4–5 Focused Meetings Bridget was the first Meetings Secretary to run varies from place to place, legislation usually each year. The intention of the Meetings a Meeting with no abstracts in Seville in 2003. requires that there is an agreed process for Committee, chaired by Bridget, was to provide Bridget excelled in presenting the minutes of the redundancy procedure. All of this is an an improved international forum for the our Scientific Meetings at Society dinners, and I additional burden for science to carry and was dissemination of scientific findings and, in doing recall an extremely entertaining after-dinner not adequately foreseen and certainly not so, to raise the profile of The Society. As you speech in the University of Cork, where a discussed forcibly with our policy makers will know, the restructuring of our Meetings has number of Members and guests found it almost when there was a window for influencing the led to a pre-meeting vetting of abstracts and impossible to suppress their laughter...was this legislation. Of course, I don’t for a moment replaced the previous system of voting at due only to a loss of audio support or the fire think that it was at all appropriate in the past Scientific Meetings. Amongst the highlights of alarm? Bridget will be stepping down as to have scientists on rolling 3 year contracts Bridget's tenure was her key involvement in Meetings Secretary in July 2006, and on behalf for decades, and am glad that those days initiating the discussions that resulted in The of the Executive, Council and membership, I Society hosting the first joint meeting with the have gone. However, with the right would like to express our thanks for her Federation of European Physiological Societies arguments made at the right time it might invaluable contributions to The Society. (FEPS) at the University of Bristol in July 2005. have been possible to have obtained Although we will miss Bridget's humour and At the Bristol Meeting, Lord Sainsbury of friendship, I know that she will retain a close agreement that ‘training’ for ‘postdocs’ could Turville (Parliamentary Under-Secretary of last for up to 6 years. Making such arguments association with The Society. Prem Kumar has State for Science and Innovation), met with the been nominated by Council as the Meetings is the role of the BSF. Executive and spent an afternoon discussing Secretary Elect and is currently liaising closely science with poster presenters. Bridget also with Bridget to plan meetings for 2007–2008. There is good news currently coming from introduced the novel concept of a Public Brussels. I am sure now that there will be a Lecture into the programme, and this practice Giovanni Mann

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final considered article, as near perfect as is practicable. So it’s only fair that it should be Ann Silver subject to greater rigour in the review process. As Chairman of the Executive Committee, But why does this process have to be so often and on behalf of The Society, I would like accompanied by verbal heavy artillery? And as for the treatment of grant proposals…don’t to thank Ann Silver for her enthusiastic even go there. The phrase ‘give ‘em both chairmanship of the Benevolent Fund for Oi, ref! barrels’ comes to mind. the past 4 years. Ann has ensured that The Physiological Society has been able On the whole, scientists are a reasonably I‘m sure everyone reading this (apart from to support many individuals. polite bunch. Of course, there are exceptions those elite scientific beings who are so brilliant but, in general, a fairly high standard of they are never on the sharp end of all this) will In a past issue of Physiology News (56, courtesy is the norm. So, for example, in the have their own examples of referees’ 40), Ann reviewed the mission of the Ben course of the average ‘free communication’ comments indicating ignorance, prejudice or Fund, which was established in 1976 session at a conference – even when the just downright disdain, so I won’t bore you when the Council was asked if The content of a paper leaves something to be with any of my personal collection. But why do Society could help the widow of a young desired – overt rudeness or hostility is pretty the reviewers of papers and grants feel moved Member whose son had special needs. rare. Unless, that is, you are disastrously to provide this kind of comment and criticism? Itself a charity, The Society may not give unlucky, or are presenting your work to the away its money but it can support the British Pharmacological Society. As a PhD The most obvious answer (apart from the administration of the Ben Fund. This student, I used to view the 5 minute question criticism being well-deserved) is that the means that all donations to the Fund can session with great trepidation, not to say refereeing of papers and grants is still be used to help what the Trust Deed calls dread. The weaknesses of my presentation, predominantly anonymous. The pros and cons those in ‘necessitous circumstances’ obvious even to myself, were certain to induce of anonymous refereeing have been broached whose work, at any level, has contributed a verbal firestorm. But pretty soon I realised many times in recent years, including in these to the ‘advancement of physiology’. I am that the worse outcome was likely to be a pages, and I do not propose to drag the complete lack of questions, leaving the subject up again. However, I still feel certain Ann would appreciate Members hapless speaker – me – in a state of personally that an ‘open’ system of refereeing making a donation using the forms suspended animation and creating an all­ would encourage reviewers to be more available on The Society's web site or round feeling of anticlimax. Questions from accurate and objective – and probably more from the London Administration Office. the audience were welcome, and the vast helpful and courteous. majority of comments – even those hinting at At our last meeting, David Brown, FRS, shortcomings in the work – were almost Another factor in the process of refereeing is was appointed as the next Chairman of invariably non-adversarial. that it is a pretty thankless task. It seems to the Benevolent Fund. me that this is fundamentally wrong, given the Giovanni Mann So at times, often having just presented a talk influence that referees exert over the career myself or otherwise sitting in the audience, I prospects of the scientist. Refereeing a paper find myself wondering who and where are the or grant is an important process. It also takes individuals responsible for refereeing one’s a lot of time, at least if done properly. Is it The Benevolent Fund written work? The ones who produce those reasonable to expect already overworked The raffle held at the Manchester comments that indicate everything from not individuals to carry out this demanding task Meeting in April raised £36 for the actually having read what you’ve written, wilful out of the kindness of their hearts? The Benevolent Fund. The winner was misunderstanding, strong personal bias anonymity of refereeing means that even Douglas Bovell (Glasgow Caledonian through to sheer spite and nastiness? Who public recognition of their contribution is University). are those guys? denied. So, in a spirit of the ‘devil’s advocate’, why not, together with a more open system of Are they the same folk who asked you the At the Annual General Meeting of the reviewing, publish the names of the referees questions when you presented the work orally Fund on 26 April David Brown was in the paper – say in the ‘acknowledgements’ a few months ago? Or are they hidden in the elected a Trustee; he had been serving section? And why not reward those hard­ anonymous sea of silent faces, keeping quiet as a co-opted member of the working referees for their efforts in some for the time being but dreaming up something committee since the AGM in 2005. tangible way (for example, book/record nasty for possible future use? Or are they He was also elected Chairman in tokens, travel vouchers, free electronic access somewhere else altogether, more interested in place of Ann Silver who had to the journal etc.)? Academics are renowned what’s going on in another lecture theatre, or completed her 6 year term on the for their cheap labour compared to, say, a in the bar? Committee of Trustees. solicitor who might expect to be paid £100­ 200 per hour for providing comparable skills, Obviously the content of a full published paper The Benevolent Fund will be holding is considerably more important than that of an but some tangible token of their efforts could a fund-raising raffle at the Main oral communication. The latter is often a form only be for the good. Meeting at UCL. The prize will be a of a ‘try-out’ of novel data, and probably isn’t choice between a Fortnum and going to get published anywhere prominent, In case anyone’s listening, mine’s a pint (real except maybe in abstract form. A full ale preferred). Or three. Mason’s food hamper or a case of published paper, on the other hand, Laithwaite’s wines. represents (or at least should represent) the Len Best

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David Greenfield In 1948, at the age of 31, Greenfield 20th century in the UK. There were of was appointed to the Dunville Chair of course, problems. Nottingham had been 1917–2005 Physiology at Queen's University of chosen partly because it was in a Belfast in succession to Henry Barcroft seriously under-doctored part of the who had pioneered research into the country and was in urgent need of a human peripheral circulation. The large new hospital, which would serve Department attracted many brilliant as the main teaching hospital for the young medical scientists. Greenfield school. Unfortunately, legal problems was always able to get the best out of over site acquisition delayed the start of staff and to inspire great loyalty. Of the building of the hospital until 1971, those whose science he nurtured during and integral with it the accommodation the 16 years of his tenure of the Chair, for the medical school. However, the eight subsequently occupied chairs and, school was committed to accept of these, six became Deans of medical students in 1970, so temporary schools, and one was twice a Vice accommodation had to be used, and in Chancellor. One became President of the early years all clinical teaching was the American Heart Association. An in existing hospitals with inadequate intercalated honours degree in space and facilities. It needed David Greenfield, who died on 17 physiology was started, and the enthusiasm, commitment, confidence November 2005, had been the graduates have held important posts. and improvisation to start the school in Foundation Dean of the Faculty of Greenfield and his colleagues laid the such circumstances. Fortunately, these Medicine and Professor of Physiology foundations for our understanding of qualities were to be found among the at the University of Nottingham for 15 the neural control of the peripheral doctors already in Nottingham and a years from 1966-1981. He was Dean of circulation. Modern treatments of team of young enthusiastic foundation the first new British medical school of vascular diseases are derived from this professors who would develop a the 20th century. Greenfield was also a fundamental work. It is a measure of medical school with a different distinguished physiologist. He had held his own and his staff’s enthusiasm that approach. The students, when they the Dunville Chair of Physiology at they were constantly presenting papers arrived in 1970, were exposed to Queen’s University Belfast (1948-1964) building and demolishing hypotheses at patients from the first week of training and the Chair of Physiology at St Society Meetings in London at a time and an intercalated degree was Mary’s Hospital Medical School. He is when the journey from Belfast entailed established for all. Greenfield was re­ remembered for his gentle but firm a sea crossing and long train journey. elected Dean for a succession of 3 year determination, generous open­ terms until he retired in 1981. The new mindedness and rigorous attention to During 1963-64 he worked in Julius school was by then firmly established detail. Comroe's department in the San with an annual entry of 130, and the Francisco Medical Centre, University University Hospital and Medical Born in Wallingford in 1917, David of California. He invented and School buildings were complete, Greenfield was educated at Poo!e developed a technique for testing though not yet fully commissioned. Grammar School and St Mary's cardiovascular reflex function (lower Hospital Medical School. He is body negative pressure) which became When Greenfield retired, the Medical believed to be the first student at St used extensively by NASA. This work School was firmly established in its Mary's to take an intercalated BSc was of great importance for our permanent accommodation and the Honours degree. He qualified in understanding of how the human dream of 1966 had become the Queen's medicine in 1940. After resident circulation can withstand first the Medical Centre, which was opened by appointments, including that of House accleration and then the weightlessness the Queen in 1977. This was his Physician to Sir George Pickering, he of space travel. Greenfield has never outstanding achievement: it proved to joined the Department of Physiology at received the credit for this that he be a notable international success, both St Mary's as a junior lecturer. He told deserved. clinically and academicaly. To mark his colleagues later that he was greatly involvement, the medical library was influenced by A St G Huggett and G W In 1964 Greenfield returned to St named The Greenfield Library. Three Pickering. There he studied the Mary's as professor, but plans were hundred and eighty five Nottingham circulation in human limbs, and developing to start a new medical doctors had qualified, the local clinical collaborated in work on the school in Nottingham. Sir George services had improved out of all haemodynamics of the foetal Pickering became the chairman of the recognition and a new medical circulation. His research on the effects University's Medical School Advisory curriculum copied by several other of acceleration and enhanced gravity on Committee. In 1966 David was medical schools, established. the circulation led to an association appointed to be Dean of the new with the Institute of Aviation Medicine school. It was a unique opportunity. It Greenfield was a member of the at Farnborough. was the first new medical school of the editorial boards of most of the

Physiology News | No. 63 | Summer 2006 | www.physoc.org 48 PN OBITUARIES cardiovascular journals, a member of The 16 years David spent at Queen’s were David, and many of those influenced by him, the MRC until 1977, then medical vintage years, not just for him, the medical were to have considerable influence on member of the University Grants school and the University, but also for the medical education and its management. Committee and Chairman of its interest in physiology and medical science it David himself introduced many new ideas as Medical Subcommittee. He was also a generated in the minds of young medical Foundation Dean in the new medical school member of the General Medical students and graduates. The upsurge of at Nottingham. He also played an important Council. local interest in physiology led to a flowering part in the foundation of the new medical of the Department. A flourishing BSc schools at the Chinese University of Hong He travelled widely, even well into Honours School was established. Many Kong, the Sultan Qaboos University in Oman retirement, advising many universities medical graduates coming to the department and the University of Southern Rhodesia planning new medical schools, most for research experience were so attracted by (now Zimbabwe) in Salisbury (now Harare). notably the Chinese University of Hong its intellectual excitement and rigour that they One of his lecturers, John Shepherd, who Kong, the University of Kuwait and the changed plans and embarked on a career in joined the staff of the Mayo Clinic, was Sultan Qaboos University, Oman. He academic physiology. selected as the Foundation Dean of the new was a member of the Hong Kong undergraduate medical school that was set University and Polytechnic Grants One of the main reasons for this was David’s up there. He has had a profound effect on Committee. charisma. Though personally modest, the academic development of the Clinic. unselfish and self-deprecating, he was Another of David’s lecturers, Bob Whelan, He was made CBE in 1977, followed intensely loyal and supportive to his juniors was appointed to the Chair of Physiology in the University of Adelaide in Australia and by the Order of St John in 1978 and the and was indefatigable in encouraging and subsequently was elected Dean of its Order of Sultan Qaboos in 1986. He helping them to reach their full potential. He Medical Faculty. Later he was appointed held the Honorary DSc from The built up their self-confidence, stimulated them to think independently, and delegated Vice-Chancellor at the universities of Western Queen's University, Belfast and the responsibility and credit to them whenever Australia at Perth and finally Liverpool in the Honorary LLD of the University of possible. He inculcated a family ethos in the UK. As with David, his experience and Nottingham. In 1987 he was elected an department that bound its members together integrity made him a much sought-after Honorary Member of The Physiological in common purpose, so carrying on a member of local and national committees. Society. Barcroft tradition. His wife Peggy was particularly helpful in fostering this ethos One of David’s MRC Fellows, Bob Coles, David Greenfield married Peggy Duane through the motherly interest she took in all was elected Clinical Dean at the University of in 1943, when she was theatre sister to the members of staff and their families. She Bristol. Another of his lecturers, Darty Mr Tom Holmes Sellors at Harefield was a wonderful entertainer. The friendships Glover, went to Australia to head Physiology Hospital, and she gave David unfailing and loyalties built up during those years at the University of New South Wales and support until her death in 1999. Their lasted for life. It was an ideal environment was elected Dean of its Medical School in son, Peter, is a computer scientist, and for young academics to see what it takes to Sydney for several successive terms. In their daughter, Catherine, a poet and run happy and successful departments. Belfast, Gary Love, Robin Shanks and I, all author. There are two grandchildren. Many of those trained there later reproduced former BSc students and lecturers in the a similar style in departments in other parts Department, were elected Dean of the Robert Graham of the world. Medical School at Queen’s over a 15 year David Banks period. All of these would happily Peter Fentem With David’s philosophy, anyone could do acknowledge how much David’s influence had moulded their outlook and career...The Adapted from The Guardian, 15 December anything if they gave it sufficient thought and collective contribution they made to medical 2005, with permission. Copyright Guardian effort. No problem was insuperable. He will Newspapers Limited 2005 be remembered for his integrity, courtesy, education over the years was considerable. enthusiasm, common sense and determination underpinned by an enormous David’s modesty and habit of self­ deprecation may have resulted in his true Personal recollections of David capacity for hard work. He had excellent manual skills in the workshop and drawing national and international contribution to Greenfield’s years in Belfast office; the equipment he made was known for medical education not receiving all the recognition it deserved. His style was to give In 1948, when David Greenfield arrived in its ingenuity, simplicity and robustness and credit for his achievements to others while Belfast as a young man to succeed Henry his diagrams were famous for their clarity. down-playing his own role. Self­ Barcroft in the Chair of Physiology, I was a advertisement and self-promotion were preclinical medical student. He so impressed Scientifically, the Greenfield years at Queen’s completely foreign to him. Perhaps only me by his enthusiasm for his subject that, were very productive. He continued Barcroft’s those colleagues who knew him well when the opportunity to take an intercalated interests in the human peripheral circulation recognized fully his truly remarkable qualities. BSc presented, physiology seemed the and thereby maintained the department’s Such qualities are in short supply today and obvious choice. Thus began one of the most international reputation. The standard test for will be sadly missed. formative year in my life and I remain assessing the circulatory reflexes of everlastingly grateful for the way it opened astronauts is based on Greenfield’s work on Ian C Roddie my mind. the effects of lower body suction on man Honorary Member, The Physiological Society

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Autar Paintal Edinburgh from Oxford). take his electrophysiological equipment there but as a penalty for not returning 1925–2004 His research uncovered many aspects to the parent institution, as required by of innervation through the vagus nerve the terms and conditions of his award, of the structures within the body cavity, he was not allowed to take his rig to especially perhaps through his work on Kanpur. Some believe they were not the heart and lung receptors. Some going to be seen as supporting the would say his discovery in 1955 of the Indian army. So he had to build J receptor in the lungs was his most everything again from scratch, in spite important contribution. of the lack of parts and the absence of funds for importing anything from I am not so sure for it is arguable that overseas: only someone with his fierce the most far reaching and most widely determination to get on with his applied discovery he made was in research and sheer tenacity in Edinburgh (1951–1953) where he overcoming endless bureaucratic showed that one could record from hurdles throughout his career in India nerve fibres, and dissect them under A personal memoir could have done it. liquid paraffin. This technique, so Autar Paintal was one of the most simple in conception, has been taken up Maybe the association with a military distinguished physiologists to emerge and used by neurophysiologists around institution had advantages for him, such from India, probably the most the world. It immediately superseded was his skill, for he would have had distinguished. The internet will give frequent irrigation of the nerve with access to military surplus apparatus. 236 references to him in <0.31 sec. saline as in the early work of Adrian or Think of those radar sets, amplifiers There one may find detailed accounts the steam-box in which tissues were and power supplies, etc. In London at of his work by a wide range of kept to prevent drying of the nerves. this time physiologists and other admirers. With that original apparatus, dissection scientists scoured the army surplus and was very difficult. Paintal’s advance second hand stores in Lisle Street, Aside from his outstanding excellence changed the way we all did Soho, in the same way. and incisiveness as a physiologist, it experiments on nerve and made those was his grace and ever youthful sense experiments so very much easier. I came to know Autar Paintal through of fun and mischief that endeared him Furthermore, the method allowed much The Physiological Society and we met to so many friends around the world. finer dissection than would otherwise often in the UK and from time to time The well chosen photograph of him on be possible, for the nerve strands do not in India. His distinction never obtruded the Vallabhbhai Patel Chest Institute dry out. Thus smaller and smaller nerve in his interactions with colleagues old web site where he worked from 1964 fibres could be studied. and new such was his openness. just about shows that twinkle in the eye, and his ‘love of fun and mischief’. Another of Paintal's innovations was to He was politically incorrect as we It is reproduced here. There is the man short-circuit David Whitteridge's very would now term it, and in some ways so many of us admired. He would have elaborate indirect ways of localizing politically incompetent, though he rose been delighted to find himself on the cardiac/pulmonary vagal afferent to the highest levels in Indian science, internet and especially to find that he endings in the anaesthetised animal by maybe because he was so was available on e-Bay in a list that fearlessly opening the chest and straightforward. His sense of fun and includes ‘new & used electronics, prodding about the heart and lungs with his political incorrectness showed itself apparel, collectibles…autar paintal dsg a glass rod. A fine example of direct after a Physiological Society dinner in accountants…holo holo…’ and so on. observation and experiment. Magdalene College, Oxford where he Especially ‘holo holo’ would have gave the vote of thanks. He told how appealed, an item closely listed near to With his PhD, he returned to India; but difficult it was to do science in India, him for sale. Similarly he is often to be not, as expected, to Lucknow and similar strictures apply throughout found as an ‘adobe acrobat’ which University – where he had obtained his the world. Monday was the day after would have provoked more glee. MD (or MB). He would not go back to the weekend, Tuesday would be a the Lucknow Medical College religious celebration day, Wednesday With the award of a Rockefeller Physiology Department because he did one played games in the afternoon, Fellowship, Autar Paintal arrived in not approve of their employment Thursday was a Saint’s day, and on Edinburgh in 1951 to work for a PhD policies, but instead took a research Friday no self respecting Indian like under David Whitteridge. He spent position in Kanpur, at the Indian Army himself could possibly do an several months first building an medical centre. experiment with the weekend electrophysiological rig with ex-army imminent. Of course, it was not like electrical parts (with circuits from CJ This failure to return to Lucknow got that, and certainly not for him. He was Dickinson's book and guidance from him into trouble with the Rockefeller devoted to science and pursued his 'Jock' Austin – both brought by DW to Foundation: they had allowed him to subject with absolute integrity, he did

Physiology News | No. 63 | Summer 2006 | www.physoc.org 50 PN OBITUARIES/FEATURE not countenance dissembling and was scientist, interested in advancing (Nicol et al. continued from p. 33) always straightforward. This quality no knowledge, pursuing investigations doubt occasionally led to trouble for with absolute integrity to the data, representative of the systems studied. him. never fearing to criticise what he Equally important, cognitive brain considered to be unworthy, and thus research often requires highly trained Many years later, as head of the Indian upholding the highest ethical standards. animals. By maximising the data MRC, he made the headlines when, obtained from each animal, we reduce aiming to prevent the spread of AIDS Certainly he was widely admired and the number of animals needed – a duty to the subcontinent, he strongly urged held in great affection by many of any bioscientist. Indian women to avoid making love to physiologists and there are many who foreigners. This view was not widely would have more stories to tell than In summary, MEA techniques alone do applauded. me. Many knew him better, going back not guarantee successfully to the Edinburgh days, but perhaps no understanding the brain, nor do they In his role as an organiser my one enjoyed being with him more. We make other techniques obsolete. They experience of Paintal in operation was cannot imagine how much more than us are a promising supplement aiming to in India particularly at the time of the his family must miss him. bridge the gap between imaging and Physiological Congress in Delhi when single-cell studies. The brain has a he organised a meeting in Srinagar. He I am grateful to Kres Krnjevic for his comments, century old track record of encrypting knew how to make administrative especially on the Edinburgh days. its functional principles. A combined systems work. Many of us arrived at strategy incorporating behavioural, the airport in Delhi but some were Tim Biscoe imaging, single cell and MEA delayed in Mumbai (Bombay as it then Honorary Member techniques will play the central part in was). Paintal moved up and down the deciphering the code. The rest is down line quietly instructing us to make our Deceased Members to the ultimate tool – our brain. After check-in as slow as possible by The Society also reports, with regret, all, it takes a thief to catch a thief! misplacing our tickets, fumbling with the deaths of D L Ingram and JA our identifications, searching for our Bateman. Acknowledgements luggage. In short we were to behave Research in our laboratory is funded by like more than usually incompetent Doug Ingram was a Member from the BBSRC. academics. This combined with the 1963–2001 and J A Bateman joined need to have every paper stamped and The Society in 1935, making him one Alister U Nicol approved by various officials delayed of our longest serving Members. Hanno Fischer our take off to such an extent that the Andrew J Tate We hope to carry obituaries in a future Mumbai group arrived to travel with us. Keith M Kendrick issue of Physiology News. Laboratory of Behavioural & Cognitive Neuroscience, He had developed to a fine degree the The Babraham Institute, Cambridge, UK use of the technique of anger to make References progress on a problem. It is always the Calder AJ & Young AW (2005). Understanding the recognition of case that to become angry is to lose facial identity and facial expression. Nat Rev Neurosci 6, 641-651. control. However, to perform as if Symposium Horton PM, Bonny L, Nicol AU, Kendrick KM & Feng JF (2005). angry is sometimes useful in Applications of multi-variate anaylisis of variance (MANOVA) to multi­ electrode array electrophysiology data. J Neurosci Methods 146, 22­ administrative life, but the technique Involvement of interstitial cells of Cajal 41. should be used sparingly and always be in the control of smooth muscle Kendrick KM, Levy F & Keverne EB (1992). Changes in the sensory under control. Paintal was a master at excitability processing of olfactory signlas induced by birth in sheep. Science this. My wife and I were once in his 256, 833-836. office, even we feeling intimidated as Saturday 22 July 2006 Nicolelis MAL & Ribeiro S (2002). Multielectrode recordings: the next he addressed some unfortunate down at the Japanese Society for Smooth steps. Curr Opin Neurobiol 12, 602-606. the telephone. So soon as he had his Muscle Research, Okayama, Japan Laurent G (2002) Olfactory network dynamics and the coding of way, he switched in a flash to his multidimensional signals. Nat Rev Neurosci 3, 884-895. normal charming self, unruffled by the Speakers include Terumasa Komuro, Tate AJ, Fischer H, Leigh AE & Kendrick KM (2006). Behavioural and previous maelstrom. G David S Hirst, Hikaru Suzuki, neurophysiological evidence for face identity and face emotion processing in animals. Phil Trans R Soc Lond B (in press). Sean Ward, Kenton Sanders, His widespread international Noel McHale, Rick Lane, Rolls ET, Treves A & Tovee MJ (1997). The representational capacity recognition is typified by his of the distributed encoding of information provided by populations of Hikaru Hashitani and Gerard Sergeant neurons in primate temporal visual cortex. Exp Brain Res 114, 149­ appearance with rather a stern image in 162. Poccuцckaя Akademuя Hayk as a Full programme available at Sugase Y, Yamane S, Ueno S & Kawano K (1999). Global and fine Foreign Member. information coded by single neurons in the temporal visual cortex. http://jp.physoc.org Nature 400, 869-73.

Always he was, whatever the Young MP & Yamane S (1992). Sparse population coding of faces in circumstances, an intensely serious the inferotemporal cortex. Science 256, 1327-1331.

Physiology News | No. 63 | Summer 2006 | www.physoc.org BOOK REVIEWS/ARCHIVE LAUNCH PN 51

Excitatory amino acid transmission in health and disease By Robert Balázs, Richard J Bridges and Carl W Cotman. 2006 Oxford University Press. 368 pp, £49.00 (hardback). ISBN 0-19-515002-3 This is an up to date, very well researched and well written book on glutamate and the The Wellcome Trust launch ways it contributes to signalling in the CNS party to celebrate the through a diversity of receptors. The authors have succeeded in producing a publication of The Journal of lucid, well organised and fundamental Physiology archive account of a complex area, profusely illustrated by Cheryl A Cotman.

The opening chapter gives an overview and perspective and sets the scene for the remaining 11 chapters, five of which deal with ionotropic glutamate receptors and another with metabotropic glutamate receptors. Then follows a fascinating chapter on glutamate transporters through which glutamate is sequestered into cellular compartments and which regulate its intracellular concentration. Five distinct subtypes of excitatory amino acid transporters have now been identified and their disruption may lead to glutamate excitotoxicity perhaps associated with degenerative diseases such as amyotrophic lateral sclerosis and Alzheimer’s disease. In addition, three vesicular glutamate transporters have now been identified which may regulate the vesicular content of glutamate at synapses and this leads into a chapter on the molecular organization of glutamate receptors in the postsynaptic density, which contains a ‘supramolecular complex of interacting anchoring proteins, glutamate receptors, signaling molecules, and cytoskleletal proteins’. This complex serves a wide variety of purposes allowing Under the Medical Journals Backfiles Digitisation Project the archives of 17 British journals synaptic modulation and plasticity, which connected with medicine, physiology and biochemistry are being put online at PubMed are considered in detail in the penultimate Central. The Journal of Physiology is part of this project. Representatives of the various chapter of the book. The final chapter deals journals gathered at the Wellcome Trust’s headquarters in London for a launch party on 11 with the roles of glutamate and glutamate May. The project is a joint venture of the Wellcome Trust, the Joint Information Systems receptors in neurological diseases. Committee and the National Library of Medicine (NLM) in Bethesda, Maryland, whose National Center for Biotechnology Information (NCBI) runs PubMed Central. The book is readable, well-produced and The photos show: Top (left): William Large (centre, Editor-in-Chief of The Journal of Physiology) with Carol should appeal to everyone, from advanced Huxley (Managing Editor) and Jonathan Goodchild (Senior Production Editor); Top (right): Mark Walport (Director undergraduates upwards, who has an of the Wellcome Trust); Centre (left): Robert Kiley (Wellcome Library); Centre (right): Prem Kumar (Deputy Editor­ in-Chief of The Journal of Physiology) with Giovanni Mann (Chairman of The Society’s Executive Committee); interest in this area. Above (left): Carol Myers (NCBI), Martha Fishel (NLM); Above (right): Tilli Tansey (Wellcome Institute for the History of Medicine). Bill Winlow See also A century (and more) of physiology now online (p. 40).

Physiology News | No. 63 | Summer 2006 | www.physoc.org 52 PN NOTICEBOARD

SOCIETY MEETINGS FEDERATION OF EUROPEAN Uwe Proske, a For more information visit The Society’s member of the NEUROSCIENCE SOCIETIES Editorial Board of web site at http://www.physoc.org Vienna, Austria The Journal of 2006 8–12 July 2006 Physiology, pictured http://www.fens.org University College London, UK earlier this year at a symposium to mark 5–7 July his retirement from Physiological Society Main Meeting, 8th INTERNATIONAL SYMPOSIUM the Department of Annual General Meeting and Young ON NEUROBIOLOGY AND Physiology at Physiologists’ Symposium From NEUROENDOCRINOLOGY OF Monash University, molecules to behaviour. Australia. AGING Ribeirao Preto, Brazil Bregenz, Austria 27–30 August MOLECULAR TECHNIQUES FOR 23–28 July 2006 LIFE SCIENCES WORKSHOPS Joint International Meeting with the For the current list of speakers and Brazilian Physiological Society and other information visit Manipulating Nucleic Acids Young Physiologists’ Symposium. http://www.neurobiology-and­ 28 August - 1 September Heidelberg, Germany neuroendocrinology-of-aging.org or contact A five day practical workshop to 13 September Andrzej Bartke ([email protected]) or introduce participants to techniques Focused Meeting Control and Richard E Falvo ([email protected]). used in molecular biology modification of excitation-contraction investigations and to facilitate development of core molecular biology coupling in healthy and diseased BRITISH ASSOCIATION FOR muscle. skills. Cost: £870 (Standard), PSYCHOPHARMACOLOGY £740 (CPD Accredited). Charles University, Prague 10–16 September PCR Theory and Practice 21–23 September 2006 Nottingham 4-8 September 2006 International Workshop Lung function Experimental Psychopharmacology 22-26 January 2007 in health and disease. Summer School A five day course to introduce Bristol, UK http://www.bap.org.uk participants to this core technique 4–5 December covering the basics to quantitative Focused Meeting New developments in EUROPEAN COUNCIL FOR Real-time PCR. Cost: £870 stress physiology. CARDIOVASCULAR RESEARCH (Standard), £740 (CPD Accredited). 2007 (ECCR) For further information and application Glasgow, Scotland 29 September – 1 October form visit our web site: 8–12 July La Colle sur Loup, Nice, France www.caledonian.ac.uk/mtls Joint Meeting of The Physiological http://www.eccr.org or contact: Mrs J Pierotti MTLS Society, Biochemical Society and Administrator, Biological and British Pharmacological Society. IUPS Biomedical Sciences, Glasgow Bratislava, Slovakia Kyoto, Japan Caledonian University, Cowcaddens 10–14 September 27 July – 1 August 2009 Road, Glasgow G4 0BA, Scotland UK Joint Meeting of The Physiological UK Tel:: +44 (0)141 331 3243 Society, the Slovakian Physiological July 2013 Fax: +44 (0)141 331 3208 Society and FEPS. http://www.iups.org Email: [email protected]

The Xth Oxford Conference UK SPIKE TRAIN ANALYSIS WORKSHOP Modeling and control of breathing – 28 August - 8 September integration in respiratory control: University of Newcastle, UK from genes to systems A major international workshop to bring together individuals from Lake Louise, Alberta, Canada neuroscience, mathematics, computing and statistics to work on developing 19-24 September novel analytic tools. http://www.ncl.ac.uk/ion/spiketrain Further information is available at Notices for the autumn 2006 issue of http://www.ucalgary.ca/~oxford06 Physiology News should reach the or email Publications Office by 30 June. Please [email protected] send contributions to [email protected]

Physiology News | No. 63 | Summer 2006 | www.physoc.org