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Fostering Communication and Collaboration
The nihCatalyst A Publication for NIH Intramural Scientists
National Institutes of Health bOffice of the Director Volume 11, Issue 5 September-October 2003
On the Road to Crystal Clarity NIda’s New Image: Nora Volkow Solving the Mysteries of Membrane Proteins: Tracing the Pathways Multiple Choices for NIH Scientists Of the Addicted Brain by Peter Kozel by Fran Pollner Question: What's complex, wiggly, n the 22 years since she got her mysterious, not very abundant, hard M.D. degree from the National to pin down, risky, essential for life I University of Mexico, in Mexico and something NIH is getting to know better than else in the world? City, Nora Volkow has taken brain anyone A. slime molds research by storm, almost single- B. giant amoebae handedly harnessing brain imaging C. policy on stem cell research to the service of addiction research. D. membrane proteins She is uncovering the neural path- Answer: D ways to addiction—be it to stimu- lant drugs, alcohol, or food—and lthough membrane proteins were charting the potholes left in addic- once thought impossible to crys- tallize, their structures are now tion’s wake. A She has con- being published routinely and under- tributed enor- stood—at least a bit—and NIH’s intra- mously to the mural research program has something to do with this progress. According to scientific litera- from published works by NCI’s Sriram Subramaniam and colleagues in Nature Structural Biology, August 2002, and the Jourrta! of Bacteriology, ture—in words NINDS’ Joseph Mindell, NIH now has March 2003 “[one of] the strongest groups of and pictures mem- Tf.iree-dimensional density map of the and, this spring, brane protein crystallographers [and] oxalate transporter, with the 12 idealized she became a membrane protein structural biologists helices grouped into three sets and superim- posed manually on the map. One set is “first” at NIH: there is.” nearly peypendicular to the plane of the She is the first These investigators are exploring new membrane: another contains bends in the techniques for the analysis of membrane woman to be- transmembrayie region; and the third set proteins are come NIDA director since the insti- and applying new devel- contains both a bend and a cuwe in the tute was founded in 1974. opments to understand how membrane transmemhrane region [color coding visible Volkow officially took office on proteins work. Spread throughout NIH’s in the online Catalyst —
The Zen oe Parking
n response to the threatened loss of 1,700 parking gladly make sacrifices for the common good, we de- spaces at NIH because of various construction cided to develop a series of transportation alterna- I projects, I have had the privilege of chairing a tives that, for many people, would be more attractive Parking Committee consisting of a cross-section of than gridlock on campus when hundreds of drivers talented NIH staff. Committee members share the look for nonexistent parking spaces. abiding belief that life at NIH is impossible without We now have 400 parking spaces at Mid-Pike Plaza, parking. All of you have been the recipients of my with an improved shuttle schedule, and 200 new spaces sanguine—yet realistic—all-hands bulletins about the at Twinbrook for those who wish to use TranShare state of parking on our Bethesda campus. (see Now, thanks to the hard work of the committee transhar.htm>). With the new security entrances, it and the Offices of Research Facilities and Research will be easier to drop people off and turn around, Michael Gottesman Services, we have been able to delay some loss of and carpools, as always, will be encouraged by guar- parking spaces and replace others on campus. The anteed spaces on campus. For those who live nearby, bottom line is that the shortage of spaces has been safe bicycling and walking are strongly encouraged. reduced from 1,700 to about 400, and we need only To put the sacrifice we are requesting in perspec- embrace alternative transportation plans for the driv- tive, if all current NIH parkers use an alternative to ers of these 400 cars by December 2003 for a period driving to campus one day in 20 (or one working day of about one year. in 4 weeks), w'e will not have a problem. To solve the remaining problem, we are appealing to a sense of community, to the desire for clean air Who says Life Isn’t Fair? Much discussion at the and healthy exercise, and to a willingness to make Parking Committee has been devoted to issues of fair- some life- and workstyle changes that will be to our ness and other matters associated with the change in advantage in the long run. our parking situation. We have been assured that ad- Rather than reiterate the content of my recent e- equate handicapped parking will be available near all mails to the NIH community, I am taking this oppor- buildings, and the ratio of red parking spaces to gen- tunity to reveal some of the reasoning that has gone eral parking spaces will not change (as an aside, it into the current plan. should be noted that the ratio of red permits to red parking spaces is the same as the ratio of general per- Finding Middle Ground (and Not Paving It Per- mits to general parking spots). manently). As you may surmise, there is a range of Some administrative fixes that would change work opinion at NIH, rtinning along a continuum from the schedules are under consideration, but much thought utmost value of ready parking on one end to the ut- needs to go into this approach to ensure that the pro- most value of trees on the other. A few NIHers favor ductivity of our workforce is not compromised. removal of all parking (and some buildings) from the campus in favor of grass and trees; some others main- Going Bananas. There is also considerable con- tain that so long as there is a single tree standing at cern about the effect of increased pedestrian traffic NIH, looking for a parking space would be an intol- during nash hours on campus, since most replace- erable burden. But we found middle ground: Starting ment parking will be farther from buildings than cur- in late September and early October, we will be cov- rent spaces, especially in the northeast corner of the ering some of our grassy spaces with temporary gravel campus near Building 31. Although pedestrians have and stacking the parking in all suitable asphalt lots. the right-of-way at properly marked crosswalks, if they During the next 12 months, construction of two cross one-by-one, traffic backs up all over campus. new multilevel parking garages will be completed, Thus, we are now asking pedestrians to “go bananas” replacing all the spaces lost to the construction of to cross in bunches of 4 to 6, leaving time between these garages. Building 33, an underground storm- for cars to move. An alternative for some would be to water management facility, and a commercial vehicle use the new express shuttle to Buildings 31 and 10 security checkpoint. So by the end of 2004, we will from lot 41. More bicycles on campus also mean that be back to our usual blissful parking state (which, I drivers must be alert and bicyclists must follow the am proud to say, includes the lowest percentage of rules of the road. single-occupancy vehicles of any major federal or The Parking Committee will closely monitor the situ- private suburban facility). ation as spaces are lost and alternative parking comes on line. If the situation deteriorates despite voluntary Self-interested Selflessness. During a portion of efforts, then we will consider implementing alterna- the next year, there will be a shortfall of up to 400 tive plans that involve scheduled exclusion of some spaces (5 percent of our total) on the NIH campus. of our staff from parking on campus. We hope this The Parking Committee considered a variety of ap- approach will not be necessaiy and we will all work proaches to reduce the demand for parking, includ- together as outlined above to solve our parking prob- ing excluding some of our staff from on-campus park- lems. As always, please send your comments and sug- ing or restricting parking for some individuals for one gestions. or more days a week or month. —Michael Gottesman But, in keeping with the evidence that NIHers will Deputy Directorfor Intramural Research 2 September-October 2003 Cartographers Draft NIH Research Road Map by Peter Kozel efore “road map” became the will achieve, Spiegel said. He noted their current mis- catchword for a Middle East that although membrane proteins sions. B peace plan, NIH Director Elias are the targets of between 40 and Katz also talked Zerhouni used the term to describe his 60 percent of current pharmaceu- about the estab- nascent plans for redirecting NIH re- ticals, very little stnictural informa- lishment of 20 to search (see “Roadblocks, Road Maps, tion is known about this class of 30 regional Trans- and The Perfect Storm,’” 777eM//(Tt/r7- molecules (see story, page I). lational Research /ysf, January-Eebruary 2003, page 5). At The image we now have of these Centers, each pro- Elias Zerhouni the June meeting of his Advisory Com- key proteins, Spiegel said, is akin viding core sup- mittee, Zerhouni and some of his chief to a 19th-century daguerreotype, port in biostatis- cartographers presented in greater de- but what we want to produce is a mo- tics, pharmacogenetics, novel reagent tail some of the research road map’s tion picture. preparation, animal toxicity testing, and byways and smaller avenues. other services. The road map is the product of l6 From Solo The behavioral and social sciences working groups comprising more than To Ensemble Performance would be tapped to develop improved 300 NIH and outside experts and NIDCR Director Lawrence Tabak pre- means of assessing clinical outcomes, in- chaired by IC leaders. It emphasizes sented the road map’s vision of its driv- cluding quality of life measures. new tools and building blocks to ex- ers—that is, multidisciplinary and inter- To bolster clinical research and accel- erate the process for performing clinical studies, NIH would work with other federal agencies and with industry to har- monize and sim- plify many of the current regulatory burdens. Rules and pro- Peter Kozel cedures govern- ing existing na- Allen Spiegel Lawrence Tabak Stephen Katz tional and interna- plore individual molecules and whole disciplinaiy research teams. It is NIH’s tional genetic and histological reposito- systems; multidisciplinary, cross-institute responsibility to break down any cul- ries would be standardized, and infra- research; and substantial expansion of tural or administrative barriers to stnicture would also be built to increase translational research. multidisciplinary research and to iden- repository availability within the clinical tify research problems that transcend the research community. From Road Blocks focus of several ICs. The road map calls Finally, the clinical research workforce To Building Blocks for the creation of trans-NIH commit- needs to be boosted by increased train- NIDDK Director Allen Spiegel elabo- tees to address these issues. ing and by expanding current efforts rated the goals of the building-block ini- Emphasis is also placed on training throughout the ICs. tiative—to characterize and elucidate the investigators to be conversant in sev- There was a general consensus that roles of all functional elements in ge- eral fields and, especially, on the cre- these initiatives will require some nomic DNA, RNA species, proteins, car- ation of several new granting mecha- changes in NIH's approach to basic and bohydrates, lipids, and metabolites in nisms to accelerate the funding of in- clinical research and in granting mecha- multiple organisms. novative, high-risk, and multidiscip- nisms. He cited NIDDK research showing linary and interdisciplinary research. Michael Gottesman, deputy director for that glycolipids have metabolic, struc- intramural research, inclicated that the tural, and signaling properties as an Translational Research: flexibility of the intramural program example of uncovering the multiple From Cross-NIH to Cross-Country makes it an ideal place for piloting some roles that molecules may have. NIAMS Director Stephen Katz pre- road map activities and complementing Increased access to tens of thousands sented the final aspect of the road map, others. of small molecules through the creation a series of proposals designed to stream- The road map continues to receive of new molecular libraries, Spiegel said, line and expand clinical research. congressional blessing: The House of will most certainly expand the scope of Translational research would benefit Representatives allocated $45 million in pharmaceuticals, which currently target from enhancement of the clinical re- its 2004 budget to the Office of the Di- fewer than 500 gene products. search infrastructure across the coun- rector, urging NIH “to maximize the use The solving of the 3-D structure of try. This would include the better inte- of the fund to implement the ‘roadmap’ the integral membrane protein rhodop- gration and coordination of clinical re- being developed by the NIH to structure sin is a model of what the road map search networks and the expansion of its future research portfolio.” H 3 Nora Volkow continued from page 1 rector for life sciences, and chair of the identify the main skeleton of what each part of the NIAAA intramural program), medical department. She had also served addictive daig has done. although I handed over many of my on- concurrently as professor of psychiatiy going projects to my colleagues there and associate dean of the medical school Q: And your methodology? when I became NIDA director. I go there at the State University of New York, Volkow: Imaging. Most of my work has once a month for three or four days— Stony Brook. concentrated on positron emission to- it’s hard, but it’s very important to me Voe NIH Catalyst interviewed Volkow mography [PET], which shows the neu- to continue doing research. in mid-August, three and a half months rochemical changes in the brain and the We are investigating the long-term ef- into her NIH job. pharmacologic properties of dmgs in the fects of drugs and alcohol on the re- brain. ward circuits of the brain, the neuro- Q: What factors informed your deci- biological mechanisms underlying vul- sion to accept the offer to become Q: What are some of your findings? nerability to addiction, and the effects NIDA director? Volkow: I was the first to document that of expectation and context of the ef- VOLKOW: It was a very difficult deci- cocaine is toxic to the human brain. The fects of drugs of abuse. If you can un- sion. I loved my job (at Brookhaven). belief at the beginning of the ’80s was derstand this, you can manipulate the But I recognized what a great oppor- that cocaine was a benign drug. I did outcome. tunity it would be. I have been in the studies that showed areas of small cere- Another area that fascinates me, and field of drug addiction since medical bral strokes in the brains of cocaine abus- for which there is an ongoing project, school and devoted all my professional ers. Nobody believed it, and it took me is the study of the therapeutic effects of career to understanding why drugs of a long time to get published. stimulant dmgs that can also be abused, abuse promote addiction and the The first study where I documented such as methylphenidate and amphet- changes in the brain of people who be- these brain changes was in 1985; I pre- amine. Why are they addictive in some come addicted. sented my findings at the Society of contexts and therapeutic in others? The possibility of going beyond the Nuclear Medicine, and they didn’t be- More recently, about seven years ago, constraints of my own work and actu- lieve it. And when I submitted the study, I started to work on obesity as a model ally having an impact on so many people the reviewers said, “There is no evidence of an addictive disorder. and on the research of the future was that cocaine is toxic to the human brain.” very challenging. I finally got it published in 1988. (Br J Q: Will you be doing any work with My predecessor, Alan Leshner, had set Psychiatry', 152:641-648, 1988). NIDDK in the area of compulsive an example of how you can shape a I actually started working with imag- eating and obesity? field. He fought the stigmatization of ad- ing in 1981, when it was not very com- Volkow: Yes. I would like very much diction. He changed the perception that mon. I was one of the first people to to work on that initiative and I’ve met addiction is something an individual use PET technology, first for the investi- with [NIDDK director] Allen Spiegel. chooses out of weakened moral stan- gation of brain tumors, then to study I am stmck by the similarities of loss dards. Addiction is a disease, not a schizophrenia [during a psychiatry resi- of control and compulsive behavior in choice. dency at New York Liniversity from 1981 people with compulsive patterns of eat- to 1984, which involved research at ing and in people addicted to dmgs. Q: How did this approach dovetail Brookhaven] and then with cocaine and The difference, of course, is that you with your own research? alcohol [at the Liniversity of Texas Medi- need food to survive, and so the inter- VOLKOW: My own research has con- cal School in Houston, as an assistant ventions would be different. centrated on identifying the changes in professor of psychiatry and behavioral But reward mechanisms, conditioned the brains of addicted people that lead science from 1984 to 1987]. responses, environmental influences— to loss of control, at how these drugs I returned to Brookhaven in 1987 spe- (such as stress and drug availability) take away a fundamental aspect of hu- cifically to learn what happens in the these variables that are so important in man behavior—free will. brain of drug abusers and why drugs of the prevention and treatment of dmg In addiction, the volitional component abuse cause addiction. addiction—can be brought to the pre- of human behavior is very much dis- vention and treatment of obesity. turbed. You can exert some control in Q: And you went to Brookhaven be- not putting yourself in certain situations, cause there was a program? Q: Are you contemplating any other but once you are in those situations, it’s Volkow: No. I went to Brookhaven on collaborations? almost like a reflex that overtakes the condition that I could develop a pro- Volkow: Yes. We are very much inter- ability to choose. gram on drug addiction. People had not ested in collaborating with other insti- How have you got to that state? ’What yet recognized how powerful imaging tutes, including those that study the has happened to the brain? could be for the study of addiction. brain, and we are working on a docu- The other question is, why can some Brookhaven had the equipment but not ment that will allow us to integrate ar- dmgs do that? You know, very few com- yet the program. Now, 90-95 percent of eas of common research in neuro- pounds can. What are their characteris- the imaging research at Brookhaven is science. tics? in drug abuse and addiction. I am also very interested in collabo- I have been investigating multiple rating with NICHD regarding the spe- drugs of abuse in parallel—cocaine, al- Q: Do you maintain a connection cial vulnerabilty of children and ado- cohol, heroin, methamphetamine—to with Brookhaven? lescents to addiction. determine the common elements, to Volkow: Yes. I keep my laboratoiy (as I want to collaborate with NIAID be- 4 " " September — October 2003 cause dmgs of abuse are extremely important in AIDS trans- mission, both through drug injection and through inducing mental states that lead to risky behavior. Collaboration with NCI is also very important because over- coming nicotine addiction is the number-one public health intervention that will have an enormous impact on cancer. Q: Dr. Zerhouni recently appointed a 10-member steer- ing committee of NIH institute directors. You are among them and the member most newly arrived to NIH. Do you think your voice will be as strong as those who have been here longer? Volkow: It’s actually very good to bring on someone who is new, who brings a fresh perspective. We have a veiy good combination on this committee of new people and people with knowledge of the system. Q: What do you think of the suggestion in the recently released Institute of Medicine report that NIDA and NIAAA be integrated into one institute? Volkow: What’s important is not whether there are one or Fran Pollner two institutes but that we recognize that these addictive dis- T!je great-granddaughter of assassinated Russian revohitionaty orders are co-morbid, that they should be studied jointly. For Leon Trotsky ("Trotsky had two sons and two daughters; myfather example, there are very few alcoholics who don’t smoke, yet was the son of one of the daughters"), Nora Volkow grew up in the house-turned-museum in Mexico i)i which Trotsky was killed. She the animal models are mostly for alcoholism. They pertain, has three sisters—a writer an economist, and an infectious therefore, only to the 10-15 percent of alcoholics who do not diseases researcher—all of whom live in Mexico. also smoke. That gap has happened not because there are Wlyen she graduatedfrom medical school in 1981, she earned not two institutes—NIAAA and NCI—instead of one but because only a degree hut a "Natio)tal Awardfor the Outstanding Medical people have not worked together. Student" as well. She came to the United States for her residency to pursue her research career "the possibilities T. K. Li [NIAAA director] and I have had several working and because for doing this kind research in Mexico, the resources, were much meetings and are planning some brainstorming sessions to of more limited. integrate our studies on genetics and dmg and alcohol addic- — Tljoiigh the resources in the United States were excellent, she did tion nicotine, too. And we will prepare an agenda for clini- occasionally encounter discrimi)jation. In one situation, her being cal trials in these areas. Mexican was cited as a "disadvantage" when she was applyitigfor a particular postgraduate program; in another, her being a Q: How long do you think you’ll be here, and where do woman was cited as a reason she should not be consideredfor a you want to take NIDA? supewisoiy positioii. In the latter instance, more enlightened minds prevailed; in theformer, she was not admitted to the Volkow: I don’t think anybody in their right brain would take program. "But Tm flexible— not having that particularprogram the position of NIDA director if they are not committed to did not stop me, "she recalls. “My task was to understand the staying at least five years. I’d like to see NIDA use the tremen- braiti. I simply did something else to get there. dous developments in science and technology to fight drug abuse and addiction. tion sciences. Why is it that the brain of adolescents is more I’m very committed to blending basic and clinical research, sensitive to drugs? What are those developmental changes to deriving concepts from basic research and applying them that make it more sensitive? How do we introduce prevention clinically, and to taking clinical findings back to the bench. at this stage? It is not so straightfoiward, not so simple. This is a common goal for all the institutes, and at NIH we Another area of missing information is the whole area of can do it. I want to create an environment where scientists of marijuana addiction. It has been, in a way, the forgotten drug, different backgrounds and cultures blend together—physi- especially in light of the fact that it is the number-one illegal cists, mathematicians, biologists, geneticists, clinicians, chem- drug of abuse. What is the extent to which early exposure to ists. marijuana increases vulnerability? What are the effects of early exposure on learning? Q: Is there a dearth of certain disciplines within NIDA? Would you want to hire people? Q: Did your family background have any effect on your Volkow: There are areas of expertise that may not be well thinking when you were growing up? represented, but we can either hire new people or create Volkow: If your family has been persecuted and extermi- collaborations to take advantage of the expertise from other nated for a cause—my father was the only one in his family institutes. You don’t want to replicate resources. It's not nec- to survive Stalin, and my mother’s family experienced the essary for every single intramural program to have eveiything. same thing under Franco—you cannot take your life for granted. It is a privilege to be alive, a privilege to have capa- Q: Are there any research areas calling out for more NIDA bilities and education. I have had all along a sense of respon- support, any where there are huge gaps in knowledge? sibility—to give something back, to help. That may have come Volkow: There are huge gaps in knowledge in the preven- from my family. 5 Membrane Proteins continued from page 1 ever, the fundamental fea- proteins but also their study duction. tures of most of these clini- by complementary tech- “So we view the barrel as a rigid scaf- cally important targets, in- niques. fold that allows the rest of the protein cluding their structure and to be positioned across the membrane function, are poorly under- Iron Transporters to interact with components in the ex- stood. With more than 15 years tracellular space and the periplasm,” Low expression level, of experience solving the Buchanan says. large size, multimeric com- crystal structures of mem- She says such structural details are the position, and complex ar- brane proteins, Susan key to molecular understanding of trans- rangement of domains con- Buchanan, an investigator port and maybe even the design of some spire to make membrane in NIDDK’s Laboratory of new vaccines. proteins notoriously trouble- Molecular Biology, ob- some to study. Unlike cyto- serves that there are still a G-Protein-Coupled Receptors plasmic proteins, membrane Susan Buchanan “huge number" of techni- Reinhard Grisshammer, a staff sci- proteins contain very large cal developments needed entist in NIDDK’s Laboratory of Molecu- hydrophobic regions, mak- to optimize the expres- lar Biology, studies the enormous fam- ing purification difficult. sion and purification of ily of G-protein-coupled receptors The crystal structures of membrane proteins. She (GPCRs). About 1,000 GPCRs have been only a handful of mamma- and other NIH scientists identified, 300 to 400 of which are found lian membrane proteins are hammering away at throughout the body binding endog- have been solved. these needed advances. enous ligands (the remainder are In the past, scientists Buchanan focuses on chemosensory GPCRs for odors, phero- could expect to spend a de- iron transporters from mones, or tastes). cade divining the optimum several gram-negative Recognizing signaling molecules like conditions for the produc- bacterial species. Iron is epinephrine and neuropeptides, GPCRs tion, purification, and crys- necessary for bacterial “are, of course, major drug targets for tal formation for a single proliferation; if iron up- disease therapy,” Grisshammer says. But membrane protein. Reinbard Grisshammer take could be blocked, his interests lie in understanding “the ba- But the times are chang- she observes, an infection sic mechanisms of how these receptors ing. Over the past five years, scientists could be stopped in its tracks. see their ligand, how they change con- here and elsewhere have solved the “Pathogens such as Neisseria formation, and how they transfer the sig- structures of nearly 80 bacterial pumps, metiUigitidis and Yersinia pestis have nal across the membrane and lead to channels, and receptors. It now takes just completely different iron transporters activation of a heterotrimeric G-protein.” one to three years to form crystals and from [Escherichia] coli, and that's a good A crystal structure for only one mam- analyze data for some bacterial pro- thing to study structurally,” Buchanan malian GPCR is known—bovine retinal teins—just about right for a postdoc notes, also pointing out that these trans- rhodposin. Kryzysztof Palczewski’s project, Buchanan points out. porters are highly antigenic and prob- group at the University of Washington A sizeable number of NIH scientists ably good vaccine targets. took many years to solve its crystal struc- are now riding this wave of discovery. Iron transporters have other impor- ture. We NIH Catalyst intewiewed seven sci- tant qualities. Energy is required for For the receptors Grisshammer is in- entists to suwey the range and depth of transport, and it appears that many dif- terested in studying, which are much less structural membrane protein discovery ferent outer-membrane iron transport- abundant than bovine rhodopsin, “you at NIH. ers are coupled to a common inner- need to do the tedious expression, pu- membrane protein involved in energy rification, and then crystallization. And From Expression transduction. Bacterial iron transporters so we have worked for 15 years on de- To Crystal Structure are picky about their passengers, usu- veloping methods to express membrane And Protein Function ally ferrying iron bound to carrier pro- proteins, including GPCRs, in functional Before scientists can solve the struc- teins rather than lone iron atoms. In fact, form and to establish robust purification ture of a membrane protein, they have they are highly specific for proteins schemes at large scale.” to be able to express it in quantities suf- binding Ee(III). In collaboration with Joseph Shiloach ficient for their method of structural Buchanan’s first crystal structure of at NIDDK’s Biotechnology Unit, analysis. If that method is crystallogra- an outer-membrane iron transport pro- Grisshammer uses a 200-liter fermenta- phy, the quantities required are large and tein yielded not only a smaller-than-ex- tion tank to obtain receptor material of the challenges of producing these quan- pected 22-stranded transmembrane P- high quality for the regular purification tities of usually scarce molecules are barrel (all predictions at the time sug- of receptors. huge. Experts here say one of the keys gested 30 or more P-strands), but an These techniques have been honed to NIH's lead in analysis of membrane additional surprise as well—a globular to the point that he can produce 5 mg proteins is its growing savvy in expres- domain sitting inside the pore. of functional receptor molecules each sion techniques. This globular domain is thought to week. These lots are used to set up crys- Advances in the expression of mem- play roles in ligand-binding specificity tallization experiments and to develop brane proteins have made possible not and interaction with the inner-mem- specific antibodies that aid crystalliza- only crystallographic analysis of those brane protein involved in energy trans- tion. 5 September — October 2003 While scientists believe that the over- be generated in reasonable amounts, but central nervous system, glutamate recep- all stmctures of GPCR families—for ex- the yield has been low, and larger cul- tors transfer excitatoiy signals between ample, the rhoclopsin family—are simi- tures have proved less efficient for pro- cells by allowing sodium to enter a cell lar, small differences tein production in response to the binding of a molecule. in the ligand-binding than small ones. Glutamate receptors “do all kinds of regions may deter- Additionally, purifi- wonderful things in the brain,” Mayer mine how tightly cation protocols notes, indicating that his goal is to un- those ligands bind. need to be derstand how they work at the molecu- “That’s the exciting tweaked. “This is lar level. Mayer says ligand binding is bit, actually, so hav- risky business,” Xia “still quite difficult to model . . . until ing stmctures from a says. you’ve got really detailed side-chain in- few representatives There are many formation for a particular subunit.” would be quite hypotheses as to Mayer currently works on kainate re- good,” Grisshammer why high-level ex- ceptors, one of four members of the observes. pression of mem- glutamate receptor family. Biochemical brane proteins is studies indicate that the protein has four P-Glycoprotein difficult. One of domains, and it’s believed that four pro- Di Xia, an investi- them is membrane tein molecules—each with 1,000 amino gator in NCI's Labo- crowding. acids—combine to form a functional ratory of Cell Biology, focuses his crys- To address the problem, Xia is using kainate receptor. tallographic efforts on proteins respon- a novel system that places expression This large size and degree of com- sible for multidmg resistance. “Resistance of transporter genes under the control plexity make it impossible to study the is everywhere,” Di Xia says. of bacterial photosynthesis promoters to intact, functional channel. “We've re- Xia’s main focus is on P-glycoprotein take advantage of the ability of photo- sorted to protein engineering to gener- (Pgp), the product of the MDRl synthetically ate soluble con- (multidrug resistance) gene. grown bacteria to structs of interest- First cloned by Ira Pastan and Michael produce large ing bits of the Gottesman at NCI in the mid-1980s, Pgp amounts of fresh molecule,” Mayer belongs to a family of transporters that membranes. says. includes the gene for cystic fibrosis. Pgp While develop- The bit on is especially important in cancer man- ing innovative which his lab is agement: Once selected in one antican- techniques for ex- currently focused cer drug, cells express the Pgp gene and pressing human is the ligand- pump many clinically important drugs, Pgp, Xia is also binding core. including anticancer agents and HIV pro- working on bacte- This domain de- tease inhibitors, out of the cell. This ATP- rial and yeast Pgp termines subtype Peter Kozel requiring pumping action reduces the homolog proteins. selectivity for Mark Mayer intracellular concentration of drugs be- He hypothesizes substrates. “The low their effective dose. that Pgps from all three groups will have receptors that have functional roles in Because cancer cells appear much similar structures and mechanisms of the brain are strikingly diverse,” Mayer more sensitive to the function of Pgp action. says. “That's due to small numbers of than do normal cells, drugs designed to Bacterial and yeast proteins are easier amino acid substitutions having very, block Pgp could be extremely useful in to study because cranking up expres- very dramatic effects on selectivity.” the clinic, Xia remarks. (According to CA: sion of native Pgp is easier than modi- The ligand-binding core can be A Cancer Journal for ClUiicians, in 2001, fying the yield of human transgenes. thought of as a clamshell, with the ion there were more than 1.2 million new Xia’s group can now purify 200 mg of channel separating the two halves of the cancer cases in the United States and protein using NIDDK’s 200-liter fermen- shell. Linkers were designed to excise more than half a million deaths, most tation tank—sufficient to begin grow- the membrane-spanning region and tie the consequence of chemoresistance.) ing crystals. With crystals in hand, Xia the two halves together. The product is How Pgp captures and removes di- will test his hypothesis by determining a soluble protein that still binds ligand, verse dmgs is not known. Xia’s lab is the similarity of bacterial and yeast Pgps implying that the structure has not been pursuing this by seeking to understand perturbed. how the protein works at an atomic Brain Receptors This approach works because “these level. Mark Mayer, head of NICHD’s Sec- multidomain proteins have quite clear In collaboration with Suresh tion on Neurophysiology and Biophys- domain boundaries,” Mayer says. “One Ambudkar at NCI, Xia began by express- ics, only recently tried his hand as a crys- can excise the portion of the molecule ing and purifying human Pgp. Several tallographer. Trained as a neuroscien- that is selective for the ligands.” common expression systems lack the tist, he has been studying the functional The engineered eukaryotic protein can cellular machinery to produce, modify, properties of glutamate receptors for be expressed in bacteria. "It’s slightly and transport human Pgp to membranes more than 20 years, “way before they cumbersome juggling 12 liters” of cul- and therefore do not generate protein. were cloned.” tures in shaker flasks, but, Mayer notes, Only in insect cells could human Pgp Found throughout the mammalian “growing bacteria is really not a prob- 7 — lem.” tions that proteins belonging to at least in 1992, he began his research program His lab has expressed, purified, and one of three CIC families are located on by “opening up Stryer [Lubert Stryer, Bio- crystallized two members of a low-af- the intracellular membranes of lysos- chemistry] and looking at what was finity kainate subfamily that have “strik- omes. downstream of receptors.” ing differences in binding properties.” The development of new methods for Hurley’s lab went on to crystallize and In fact, he remarks, “the drug industry measuring ion currents from currently solve structures of important domains is very interested in these because they inaccessible places, such as lysosomes, from such classical second messengers are potential analgesic and anticonvul- is the second prong of Mindell’s research as protein kinase C, phospholipase C, sant targets.”. program. and adenyl cyclase. “The hope is to broaden the number Hurley’s lab has now moved from Chloride Channels of channels that we can actually look at looking at protein domains to tackling Joseph MindeU, an investigator in the and therefore start to look at the con- protein complexes using a structural NINDS Membrane served properties of genomics approach. Thus, instead of Transport Biophysics the family vs. things looking at individual molecules, his lab Unit, applies a range that might be spe- will now be sun^eying and comparing of methods to exam- cific to [just] one or domains and structures of whole pro- ine how chloride two. And in the tein families. channels (ClCs) meantime, we may “A lot of trafficking is carried out not work. Broadly ex- learn something by single domains but by complexes that pressed in tissues, about how the fold together,” he observes. He is work- ClCs play roles as di- channels in lysos- ing in collaboration with Juan verse as salt trans- omes work.” Bonifacino’s group in NICHD to study port, brain function, sorting into endosomal pathways. muscle activity, and New Approaches Hurley has also begun to look at other maintenance of intra- To Structure protein complexes involved in subcel- Peter Kozel cellular pH. Although the lular targeting and membrane traffick- Joseph MindeU For historical rea- structures of even ing. sons, the CIC family large membrane wasn’t well studied, but proteins can be solved Electron Microscopy that’s changing, Mindell by X-ray crystallo- Sriram Subramaniam, chief of the says. He developed a graphic techniques, Section on Biophysics at NCI’s Labora- stmcture of a CIC using these snapshots have tory of Biochemistry, is focused on de- electron microscopy key limitations beyond veloping and refining an alternative im- and is now “going back crystallization. aging modality: high-resolution electron to look at function in For example, move- microscopy. light of structure.” ment is an inherent The immediate product of a high-reso- Mindell is taking a property of functioning lution electron microscopy experiment two-pronged approach transporters, receptors, is an image of a protein or protein crys- to this goal. Little is and channels. A crystal, tal that contains three-dimensional data Peter Kozel known about which however, is a static compressed into two dimensions, much James Hurley and where—portions of structure; proteins must like an X-ray image in a computed to- bacterial CIC molecules move during be locked into one, and only one, con- mography (CT) scan. Subramaniam’s lab chloride ion transport. Mindell’s lab is formation for X-ray crystallographic uses a suite of techniques to extract the addressing this by attaching fluorophores analysis. information present in these images to to different regions of the protein. The reconstruct the structures of proteins, fluoresence characteristics of the Structural Genomics protein complexes, organelles, and even fluorophores change depending on the One critical class of molecules, amphi- whole cells. hydrophobicity of their environment. trophic membrane proteins, resides in Armed with this information, Mindell membranes only when they’ve been ac- Electron Crystallography. One of the can then create, express, and eventu- tivated. James Hurley, a senior inves- techniques being used is electron crys- ally crystallize and solve X-ray stinictures tigator in NIDDK’s Laboratory of Molecu- tallography. Membrane proteins can be for mutants “to get insight into how the lar Biology, says “a great deal of signal crystallized in the plane of lipid bilayer whole thing works,” he says. “The bac- transduction involves signaling enzymes to form two-dimensional crystals. terial system is a great system for that,” that translocate from the cytosol to mem- “The goal of electron crystallography,” he notes. “’We can make as much [CIC] branes upon activation.” Hurley’s group Subramaniam says, “is no different from as we want, and it’s easy to manipulate studies these proteins, which are in- that of X-ray crystallography. ’We want in the large quantities that we need for volved in trafficking as well as signal- to solve atomic structures.” However, these sorts of classical biochemical ex- ling. because the protein is in a lipid bilayer, periments.” Hurley’s undergraduate training was it is closer to its native environment than Mindell says that while he was focus- in particle physics. He developed his in- it is in the 3-D crystals used for X-ray ing on structure, lots of fascinating “bi- terest in signal transduction during his crystallography. ology has emerged,” including indica- doctoral training. Upon arriving at NIH This special feature provides an op- 8 . ) portunity to study physi- lular membranes. “We dis- Buchanan notes that the wide variety ologically meaningful covered that these pro- of scientists at NIH is one of its strengths. conformational changes teins form these remark- Grisshammer concurs, saying that at using electron crystallog- able assemblies inside most institutions “you have to solve the raphy. cells, which we called zip- problems by e-mail or telephone. Here, By analyzing and com- pers,” Subramaniam re- you can just go and ask for help or ad- paring dilfraction patterns counts. The observation vice or materials.” of 402 different crystals of of this novel form has led To spread NIH’s experience even fur- the proton pump the lab to propose a new ther, Grisshammer has organized a mem- bacteriorhodopsin, each picture of the mechanisms brane protein scientific interest group pattern taken at a slightly by which bacteria re- ( Student Posters Shine through a Rainy Summer by peter xozei and ceua Hooper This year's Summer Poster Day (August 7), the annual exhibition of their research by students in NIH summer research programs, attracted so many participants it had to be divided into morning and afternoon sessions. Following are but eight of the record 484 posters presented throughout the day. Angiogenesis Inhibition any kinds of tumors secrete mol- ing one of two identical twins working M ecules that promote the formation at NIH this summer, also got the sexy of new blood vessels, allowing them to research topic—stem cells. Cohen, continue their rapid growth. NIDCD in- working with NIAMS mentors Wan-Ju vestigators Zhong Chen and Carter Van Li and Rocky Tuan, cultured mesen- Waes had previously observed high lev- chymal stem cells from adult human els of hepatocyte growth factor in pa- bone marrow. tients with head and neck squamous cell His goal was to compare growth of carcinomas. Brian Worden, an HHMI the cells on the usual flat surfaces to scholar working in that lab the past year, growth on biodegradable, potentially identified Egr-1 as a potentially impor- implantable 3-D nanofibers made of tant transcription factor in the expres- poly(e)caprolactone (PCL). The re- sion of platelet-derived growth factor sults—equal or slightly superior prolif- Celia Hooper and vascular endothelial growth factor. Double Take: Seth (left) andJoshua Cohen eration on the PCL film—bode well for Continuing Worden’s work, Joshua attend the same medical school but worked in the possibility of someday implanting Cohen succeeded in elucidating the different NIH labs this summer the nanofiber disks, loaded with stem Egr-1 signaling pathway that turns on cells, as frameworks for regrowing car- NIH has been an “amazing” experience. these two angiogenic growth factors. He tilage. The cells on nanofibers also “You study and read about stuff in medi- also showed that antisense oligos to Egr- showed greater predilection for calcium cal school. Here you get to work with the 1 inhibited expression of Egr-1 in a squa- mineralization, beginning this differen- people doing that research.” He thanked mous cell carcinoma cell line. tiation into osteocytes on day 1 vs. day by name his preceptors Van Waes and This type of head and neck tumor is 4 for the monolayer. Chen—and everyone else in the lab—for often detected too late for the usual Like his twin, Cohen is a second-year teaching him how to cany out experiments. therapeutic regimen to be effective; it’s medical student at Northwestern Uni- His fate, he says, is now sealed: “I have to hoped that blocking Egr-1 will eventu- versity in Chicago. He came to NIH stay in research.” P.K. ally prove clinically useful, Cohen notes. “The Big House”—for its world-class Cohen, a second-year medical student Stem Cell Platforms research and researchers and would at Northwestern University Medical erhaps it is fitting that Seth Cohen, like to combine research with medical School in Chicago, said that working at P who declares himself the better-look- practice in his career. — C.H. A Model Receptor Bipolar Comorbidity Michael DiPrima says the most tudies in adults show that important thing he learned at S anxiety disorders are preva- NIH this summer was “how to lent in those with bipolar dis- learn.” A senior this fall in order (BPD). Anna Binstock, Carnegie Mellon’s biophysics working with NIMH’s Ellen program (Pittsburgh, Pa.), Leibenluft and Daniel Dickstein, DiPrima built upon his biophysi- sought to learn whether that cal coursework to refine a model would be the case in children for the G1uR6 glutamate recep- as well. Binstock examined tor. Upon binding this excitatory medical records of young pa- neurotransmitter, the receptor tients with BPD, collected as opens, allowing ions to enter a part of a larger, ongoing study postsynaptic neuron. of pediatric BPD. Mark Mayer’s lab in NICHD, Most of the 31 patients in the DePrima did his work, is sample displayed anxiety, a co- Celia Hooper where Celia Hooper studying the mechanisms of morbidity rate in line with the Michael DiPrima Anna Binstock ligand binding to G1uR6 based 25-90 percent range reported on X-ray crystallographic diffraction patterns (see p.8). These in adults. Moreover, children with both BPD and anxiety were patterns are translated using computer algorithms into atomic more functionally impaired, with an earlier age of BPD diagno- models—but you have to know which algorithm to use. sis and more frequent psychiatric hospitalizations. Though not DiPrima’s project was to generate a model using one soft- a focus of the study, Binstock notes that treatment of these ware package and compare it to a model his preceptor cre- patients may be tricky, because standard antianxiety agents ated using a different program. When DiPrima’s model proved may exacerbate BPD symptoms. more accurate, the student became the teacher: He spent his A sophomore at Cornell University, in Ithaca, N.Y., Binstock last week here teaching Mayer how to use the better com- plans to be a pediatric psychiatrist. She enjoyed her experience puter program! DiPrima hopes to continue teaching after gradu- here shadowing clinicians as they interacted with children and ate training in biophysics. —P.K. seeing how psychiatric diseases manifest themselves. —P.K. 10 Skptember — October 2003 Sights and Smells Signaling Surprises Rhonda Moore is a unique Enjoli Cooke points participant in the summer re- proudly to her picture- search program: She has a Ph.D. perfect Western blots. in anthropology. She was pur- The senior from St. suing postdoctoral research at John’s College in An- the M.D. Anderson Cancer Cen- napolis, Md., says the ter in Houston when she caught most important thing the neuroscience bug. she learned this sum- Working in Alan Koretsky’s mer in Peggy Zelenka’s lab in NINDS this summer, she NEI lab w'as the pa- used manganese as a contrast tience it takes to get agent to enhance the visualiza- clear data—even when tion of the rat olfactory system. they point to negative She anesthetized and intubated results. “It took two and rats, applied a manganese chlo- a half months to get ride solution to their noses, and there," she laughs. placed the animals inside an Cooke was following instalment. re- Celia Hooper MRI Manganese up on studies suggest- Rhonda Moore duces Tj relaxation time, caus- ing that epidermal Enjoli Cooke (left) and mentor ing positive contrast enhancement in tissues where it accu- growth factor (EGF) has Pegg)! Zelenka mulates. Future studies will determine which portions of different effects on ceils the olfactory bulb respond to specific odorants. at different concentrations. Using her lab’s immortalized rab- Moore notes that while this particular contrast agent may bit lens epithelial cells, she exposed the cells to a range of not be as useful in human brain studies, it is an invaluable concentrations of EGF, then looked at the response of the means of studying olfaction in knockout animals that are three key initial pathways set off by EGF signaling to see models for human disorders. Now committed to mastering whether they responded differently from one another. various imaging modalities, she says a key factor in her The beautiful theory—that the varying response of the cells selection of a graduate biology program will be the avail- to low vs. high EGF levels was due to different responses of ability of an MRI system. —P.K. the MAPK, PI3K, and PLC-y signaling pathways—is now threat- ened by an ugly fact. In Cooke’s experiment, all three path- ways responded similarly, switching on and staying on from Fishingfor Folate Effects low concentrations to high. Cooke says more experiments are needed to put the theory to rest—ancl that her enthusiasm for ating CO-3 fatty molecular biology remains intact. She wants to pursue postbac acids from fish E studies in proteomics. — C.H. can ameliorate risk factors for heart dis- ease, but levels of con- sumption of fish are Stem Cell Markers low in the United Zishuo Hu, working with Tobi States. Working with Limke and Mahendra Rao at NIAAA investigator NIA in Baltimore, has been in John Umlrau, medical pursuit of useful markers that students Sunil Patel would distinguish neuronal and Karl Dauphinais stem cells in the subventricular set out to explore zone (SVZ) of the embryonic whether folate sup- mouse brain from cells in the plementation might in- ventricular zone ( VZ) of the de- crease the absorption veloping cortex. Other scientists Celia Hooper of dietary (0-3 fatty ac- have found transcription factors Sunil Patel (left) and Karl ids, especially of the that distinguish VZ cells from Dauphinais long-chain DHA form more mature cells in the devel- spinal cord. —a possibility suggested by a correlation between red blood oping cell folate levels and plasma levels of DHA in rats. Hu wanted to see whether Using human semm samples collected for another study these markers could be used in in the lab, they observed a similar correlation in humans. developing cortex but also Celia Hooper Interestingly, high levels of folate correlated with high lev- sought extracellular markers Zishuo Hu els of DHA, but not of other co-3 or co-6 fatty acids. They are that could be used to sort cells. proposing future studies to determine whether plasma DHA Working with tissue from l6-day mouse embryos, Hu was levels rise in people who take folate supplements. foiled in his goal of distinguishing SVZ from VZ cells, but did Patel, a second-year medical student at Midwestern Uni- come up with some nice markers that could be used to dis- versity, Chicago College of Osteopathic Medicine, at tinguish SVZ from VZ stem cells jointly from more differenti- cells. preliminary to Downer’s Grove, 111., wants to become a cardiologist and ated surrounding This work pointed CD24 tackle the problem of sudden cardiac death. Being able to and PDGFRa (platelet-derived growth factor receptor-a) as transform research findings into novel ways to treat patients, non-VZ and SVZ ceil markers, and Soxl as a potential VZ he says, "is the best way to practice medicine." Dauphinais, and SVZ marker. a fourth-year medical student at the University of Miami, Hu says he greatly values this lab experience and learning will be starting an internal medicine residency and hopes to that “science can be difficult and exciting at the same time." do clinical research as an adjunct to a practice that empha- A sophomore at Brown University in Providence, R.I., he says sizes preventive care and patient education. —P.K. he is strongly leaning toward a career in research. —C.H. 11 The NIH Catalyst People Recently Tenured bed nucleus of stria terminalis that play in relapse. These stimuli Yavin Shaham received his M.A. in — dmg can be major roles in stress-induced relapse.^’’ divided broadly into two categories: 1) 1988from the Hebrew Uyiiversity, Jerusa- Over the last several years, our re- discrete dmg cues (such as a needle or lem, and hisPh.D. in 1992from the Uni- search has been extended to successful white powder) that are temporally as- formed Sewices University of the Health Sciences, Bethesda. His postdoctoral human studies on the effect of stress on sociated with the acute rewarding ef- drug craving. Also, two drug classes fects of the dmg and 2) contextual dmg training from 1992 to 1995 was at found effective in our rat studies are cues (such as a specific bar) that can Concordia University, Montreal, in the either in clinical trials for relapse pre- become predictors of drug availability, laboratory ofJane Stewart. Subsequently, vention (a-2 adrenoceptor agonists) or but are not temporally associated with he was an affiliated scientist at the Ad- in planning phases for such trials (CRF^ the acute rewarding effects of the dmg. diction Research Center, Toronto, and receptor antagonists). Many laboratories currently explore an assistant professor in the Psychology Incubation of cue-induced co- the neuronal events underlying the ef- Department at the University of Toronto. caine seeking. Cocaine addiction is fect of discrete cues on relapse. In con- He joined NIDA in 1998 as a tenure- characterized by high relapse rates after trast, little is known about the mecha- track investigator, and he is currently prolonged withdrawal periods. Gawin nisms mediating dmg context-induced the chief of the Neurobiology of Relapse and Kleber hypothesized that human ad- relapse. Section of the Behavioral Neuroscience Branch. dicts become progressively more sensi- We recently adapted a rat model (origi- tive, over the first months of nally developed by Bouton and Bolles In the Neurobiology of Re- withdrawal from cocaine, to in fear-conditioning studies) to study the lapse Section,* we use a rat drug-associated cues. Experi- effect of the drug context on relapse. model to study neuronal mental evidence for this hy- We found that in rats trained to self-ad- mechanisms that may under- pothesis, however, was not minister speedball (a mixture of heroin lie relapse to heroin, co- available. and cocaine), re-exposure to the drug- caine, and methamphet- We developed a rat model taking context reinstates drug seeking amine seeking induced by to study the effect of the co- after extinction of the self-administration stressors and drug-associ- caine withdrawal period on behavior in a different context.® ated cues. In the rat relapse cue-controlled dmg seeking. We are currently exploring neurotrans- model, we measure resump- We found a progressive en- mitters and brain sites involved in con- tion of lever-pressing behav- hancement, over two months text-induced reinstatement of dmg-seek- ior induced by drug or of withdrawal from cocaine, ing. nondmg stimuli, following training for of lever-pressing behavior after expo- intravenous drug self-administration and References sure to cocaine cues.^ These data sug- 1. Y. Shaham, Stewart, “Stress reinstates subsequent extinction of the drug-tak- J. gest that the individual is most vulner- heroin self-administration behavior in daig-free ing behavior. Our main projects are de- able to relapse provoked by dmg cues animals: an effect mimicking heroin, not with- scribed below. drawal," PsvcZwp/tartnacotog)' 119: 334—341, at time points that are well beyond the Stress-induced relapse. Human 1995. acute withdrawal phase. 2. Shalev, L. Lu, Wit, studies report that stress increases re- Y. Shaham, U. H. de J. In a collaborative study with Tsung- Stewart, “The reinstatement model of drug re- lapse to dmg use, but until recently there Ping Su and Temo Hayashi from our lapse; history, methodology and major find- has been no animal model to study the ings," Psychophannacology 168:3-20, 2003. institute, we found that the time-depen- mechanisms mediating this effect. Dur- 3. Y. Shaham, S. Erb, Stewart, “Stress-in- dent changes in cue-induced dmg seek- J. ing my postdoctoral training, Jane duced relapse to heroin and cocaine seeking ing over the first 90 days of withdrawal in rats: a review,” Brain Res. Rev. 33:13-33, Stewart and I developed a rat model to are associated with alterations in brain- 2000 . study the effect of stress on relapse in derived neurotrophic factor within com- 4. U. Shalev, J.W. Grimm, Y. Shaham, “Neu- rats. We found that exposure to mild to and cocaine: a ponents of the mesolimbic dopamine robiology of relapse heroin intermittent footshock, a common stres- review,” Pharmacol. Rev. 54:1^2, 2002. reward system.'^ sor in animal studies, reliably reinstates 5. J.W. Grimm, B.T. Hope, R.A. Wise, Y. In another collaborative study with of cocaine craving after heroin seeking after prolonged with- Shaham, “Incubation Bruce Hope from our branch, we also withdrawal,” Nature 412:141-142, 2001. drawal periods.^ found that in the brains of cocaine-ex- 6. J.W. Grimm, L. Lu, T. Hayashi, B. T. Hope, Subsequently, we and others found T.P. Su, Y. Shaham, “Time-dependent increases perienced rats, glutamate receptors in that this effect extends to other drugs of in brain-derived neurotrophic factor protein regions of the mesolimbic dopamine abuse (cocaine, nicotine, alcohol) and levels within the mesolimbic dopamine sys- systems are upregulated for up to 90 tem after withdrawal from cocaine: implica- to certain other stressors.^ We also iden- days after drug withdrawal." tions for incubation of cocaine craving,” J. tified two brain neurotransmitters—cor- We and several other laboratories are Neurosci. 25-.742-147, 2003. ticotropin-releasing factor (CRF) and 7. L. Lu, Grimm, Y. Shaham, B.T. Hope, exploring further the neuronal mecha- J.W. noradrenaline—and two brain sites—the •Molecular neuroadaptations in the accumbens nisms involved in the new phenomenon central nucleus of the amygdala and the and ventral tegmental area during the first 90 of incubation of cue-controlled cocaine days of forced abstinence from cocaine self- 85:l604- * Thesectio}} currently comprises two vis- seeking after withdrawal. administration in rats," J. Neurocbem. 1613, 2003. itiyigfellou’s, LinLu andJeyiniferBossert, The drug environment and re- 8. H.S. Crombag, Y. Shaham, “Renewal of two students. lapse to heroin and cocaine. In hu- and Jack Dempsey and drug seeking by contextual cues after pro- Shirley Liu, whose bard work, and dedi- mans, environmental stimuli associated longed extinction in rats,” Behav. Neurosci. cation are gratefully acknowledged. with drug intake play an important role 116:169-173, 2002. 12 — . . . DOE Expands Half-Life Service Is Your Research Taking Some New Isotopic Directions? adioisotopes can be used for as aluminum-26, are sold from inven- tions to research, medical, and com- tory. The final two categories, research R much more than just Southern mercial customers. These forms are blots—they are also powerful tools and commercial isotopes, have much due in late spring, when they are for the diagnosis and treatment of shorter half-lives—2.6 days to years peer-reviewed. The final list of iso- disease. But more exotic biomedical and are produced as needed. topes to be produced in a subsequent is research applications may call for Actinium-225 an interesting ex- fiscal year is made public by late sum- more exotic radionuclides. NIH sci- ample. This isotope, with a half-life of mer, and orders are accepted until the entists can now request special—and 10 days, is produced from fissile, Cold- end of September for the next fiscal more mundane—isotopes for their War legacy uranium-233. An a-emitter, year. (Scientists have until September research from the U.S. Department Ac-225 shows promise as a cancer treat- 30 to place their orders for fiscal year of Energy (DOE). ment. 2004.) Payment must be made 30 days Some of the clinically important Although the DOE builds and main- before the start of production to cover isotopes, such as those used for tains unique facilities to produce isotopes production and isolation costs. positron emission tomography scan- such as Ac-225 that simply aren't avail- A fact sheet with contact informa- else, it ners, are so short-lived that they must able anywhere does not have the tion and lists of isotopes available in be produced on the NIH campus. funds to produce isotopes that do not EY2004 can be downloaded from The DOE produces and distributes have buyers. The agency has thus es- A New Kind ofNAPster Don’t Get Stuck Suggestionsfor NIH Organizational Changes, Without Your PIN Compliments of the National Academies Press TI) eginning October 1, 2003, A nyone interested in the full text of the latest outside effort to evaluate the JZ) NIH employees will use Em- ./^structure of NIH entitled Enhancing the Vitality the National Insti- — of ployee Express, an automated sys- tutes Health: Organizational Change to Meet New Challenges—may now of tem that provides access 24/7, for “read it free online” at management of the following trans- 13 The NIH Catalyst The Good News: Finnish Immunologist: People Want To Keep Their NIH Jobs New Fogarty Scholar IH’s newest Fogarty Scholar, N Helgi Valdimarsson, will be on campus for three months each fall from 2003 to 2007. Professor and chair of immunol- s/llo; A 7£ ogy at the University of Iceland, l.OWEST I WEAPON I A Reykjavik, Valdimarrson’s early re- BIDDER EDFRAL OF ,\1ASS search led to the first report—pub- lished in Nature in 1975—of what is now known as natural killer cell activity. He went on to explore the immunological underpinnings SALK of such condi- LOWEST BIDDER tions as rheu- matoid arthritis and psoriasis, elucidating the mechanisms of phagocytosis, T-cell recogni- Helgi Valdimairson tion of anti- gens, the MHC system, and other immune system pathways. Jacqiie Ballard (foreground, right), president of the NIH chapter ofBlacks in Government, His hypotheses, first, that psoria- addresses a noontime rally July 1 7protesting A-76. A sea ofsigns proclaim such sentiments sis is a T-cell-mediated disease and, as "A-76 A Weapon ofMass Distraction" and "A-76 Bush, Not Federal Workers." second, that it is triggered by strep- tococcal superantigens, changed the isplays at NIH are usually in the Blacks in Government (BIG), the event way the scientific community views D form of scientific poster presenta- was billed as an informational rally. psoriasis. His studies have extended tions, and protests at NIH (rare) are usu- Jacque Ballard, NCI, president of the to identifying candidate gene loci ally conducted by outside groups. But BIG chapter, noted that for 2002, 466 for susceptibility to psoriasis and the on July 17, the demonstrators were NIH vacant positions had simply been con- development of effective immuno- employees, and the poster boards on tracted out, affecting no current NIH suppressive therapies. display decried the A-76 competitive employees. The complement of 2003 In continuing work, Valdimarsson outsourcing process. jobs on the line, however, affects 1,000 intends to focus on identifying al- A month after a Town Hall meeting NIH employees in grants technical sup- leles predisposing to psoriasis and that had been dominated by employee port and real estate property manage- autoepitopes recognized by psoria- concerns about A-76 (see ’‘Bread-and- ment—“mostly blue-collar workers, mi- sis-causing T cells. He will also ex- Butter Dominates Director’s Town Hall norities, and women," Ballard said in a plore the use of psoriasis as a model ” Meeting, Tlje NIH Catalyst, July-August speech. More of the same is slated for for autoimmune diseases in general. 2003, 3), the demonstration revealed 2004 and 2005, she said. P- "While at NIH, he will be hosted continuing dismay over the speed with As the Catalyst to press, the went by Snorri Thorgeirsson, NCI, and which competitive outsourcing is being House had passed an amendment to an "Warren Strober, NIAID, and inter- carried out and the potential loss of jobs. appropriations bill, submitted by Chris act especially with the Immunology Organized by the local branch of the Van Hollen (D-Md.), that would thwart and Cytokine Interest Groups. American Federation of Government the administration’s A-76 plans. Sen- The Strober, chief of the mucosal im- Employees and the NIH chapter of ate had not addressed the issue. munity section, anticipates talking with Valdimarsson about the immu- nological similarities between pso- riasis and Crohn’s disease—espe- cially regarding the proximity on chromosome 16 of loci associated with the two conditions and the he online version of TJoe NIH Catalyst can be found at potential role of the gene product T 14 Ifs Not Too Late! ith a $5 late fee, late registra- Principles and Practice The course objectives are: tion for the FAES (Foundation the basic W To understand epide- Clinical Research for Advanced Education in the Sci- Of miologic methods involved in clini- ences) Graduate School at NIH is he deadline for registering for the cal research. being accepted through October 9. 2003-2004 course on “Introduc- To be grounded in the principles T $10 will enable late registration tion to the Principles and Practice of of clinical research ethics and the le- through October 24—but that is the Clinical Research” is October 3. The gal issues and regulations involved in last possible day. human subjects research, including course runs from October 20, 2003, The FAES 2003-04 course catalog to February 24, 2004, and is held on the role of IRBs in clinical research. is available on line as a PDF file at To become familiar with the prin- the NIH campus Monday and Tues- 15 The NIH Catalyst Catalytic Bubbling Biluons: A Yeast Experiment Reactions? or anything to grow, it needs the right conditions. In this experiment, F we’re going to find out how to wake up something that looks like sand—dry yeast. f you have a photo or Just looking at this stuff, you’d never think it could be used to make I other graphic that reflects bread, or wine, or pizza. But you’ll soon see that you can make this dull, an aspect of life at NIH dry sandpile just bubble over with enthusiasm! (including laboratory life) First, it might be a good idea to get a grown-up to help you gather the or a quotation that scien- ingredients and equipment you’ll need (and don’t be surprised if the grown- tists might appreciate that up gets curious and wants to stick around to see what happens). would be fit to print in the You will need: space to the right, why not A package of dry active yeast send it to us via e-mail: Four small glasses (cordial size would be good, but test tubes would be even better) [email protected]>; A packet of sugar fax:402-4303; or mail: Warm and cold tap water Building 2, Room 2W23. Your sense of what is warm and cold is just fine. (You might also write down your observations, glance at the clock every now and then, and use some wooden chopsticks to stir.) Also, we welcome Now let’s get started. “letters to the editor” for So that you remember what to do and what is in each glass container, label the four glasses (with a publication and your Post-It) with the following: 1) Yeast-warm-sugar. 2) Yeast-cold-sugar. 3) Yeast-warm. 4)Yeast-cold. Step 1: Open the package of yeast and pour it on a plate. There’s not there at all reactions to anything on much —and we’re going to divide that little bit into quarters, too! (Not much? We’ll see.) Shake the dry yeast it the Catalyst pages. so covers the plate, then divide it into quarters (an index card is a good tool for this). Put each quarter in its own glass. Step 2: Into the two glasses that are marked to have sugar, sprinkle enough sugar to cover the surface In Future Issues... of the yeast—a healthy “pinch." Step 3: Now for warm and cold. Fill a separate container with warm tap water that feels like bathwater ^ IRP Research and another separate container with cold tap water that feels like a cold soda. Then pour the warm water Roundup into the two yeast glasses that are marked for warm, and pour the cold water into the two yeast glasses that are marked for cold (look at the label to make sure, and pour the water almost to the top). I Research Festival Step 4: Stir. Use separate stirrers for the glasses with and without sugar. And CC’s 50th Step 5: Wait and wonder—but you won't have to wait long at all to see something happening. So, what is it that makes dry, dull yeast wake up and bubble over? When you decide what that is, see Phoenix Tales if you can get all four yeast samples to behave like that. —-Jennifer White For more information aboutyeast (and action videosj, see Tfje NIH Catalyst is pub- PlIBlJSHER Scientific Editor Editorial Advisory Board lished bi-monthly for and by Michael Gottesman Celia Hooper Jorge Carrasquillo, CC the intramural scientists at Deputy Director David Davies, NIDDK NIH. Address correspon- for Intramural Research, OD Managing Editor Dale Graham, CIT dence to Building 2, Room Fran Pollner Hynda Kleinman, NIDCR 2W23, NIH, Bethesda, MD Editors Elise Kohn, NCI 20892. Ph: (301) 402-1449; John I. Gallin Copy Editor Susan Leitman, CC fax: (301) 402-4303; Director, Warren Grant Magnuson Shauna Roberts Bernard Moss, NIAID e-mail: U.S. Department of Health AND Human Services FIRST-CLASS MAIL National Institutes of Health POSTAGE & FEES PAID DHHS/NIH Building 2, Room 2W23 Permit No. G-802 MSC 0235 Bethesda Maiyland 20892 Official Business Penalty for Private Use $300 0 Printed on 50% recycled conieni paper and can be recycled as office