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Characterization of the Macrostomum Lignano Ageing Phenotype

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Characterization of the Macrostomum Lignano Ageing Phenotype Characterization of the Macrostomum lignano ageing phenotype Jeroen CRUCKE Thesis submitted to obtain the degree of Master in Biology Academic year 2009/2010 Supervisor: Dr. Maxime Willems Co-Supervisor: Prof. Dr. Wouter Houthoofd Tutor: Drs. Stijn Mouton Nematology Section Characterization of the Macrostomum lignano ageing phenotype Jeroen CRUCKE Thesis submitted to obtain the degree of Master in Biology Academic year 2009/2010 Supervisor: Dr. Maxime Willems Co-Supervisor: Prof. Dr. Wouter Houthoofd Tutor: Drs. Stijn Mouton Nematology Section © May 10 Faculty of Sciences – Nematology Section All rights reserved. This thesis contains confidential information and confidential research results that are property to the UGent. The contents of this master thesis may under no circumstances be made public, nor complete or partial, without the explicit and preceding permission of the UGent representative, i.e. the supervisor. The thesis may under no circumstances be copied or duplicated in any form, unless permission granted in written form. Any violation of the confidential nature of this thesis may impose irreparable damage to the UGent. In case of a dispute that may arise within the context of this declaration, the Judicial Court of Gent only is competent to be notified. Deze masterproef bevat vertrouwelijk informatie en vertrouwelijke onderzoeksresultaten die toebehoren aan de UGent. De inhoud van de masterproef mag onder geen enkele manier publiek gemaakt worden, noch geheel noch gedeeltelijk zonder de uitdrukkelijke schriftelijke voorafgaandelijke toestemming van de UGent-vertegenwoordiger, in casu de promotor. Zo is het nemen van kopieën of het op eender welke wijze dupliceren van het eindwerk verboden, tenzij met schriftelijke toestemming. Het niet respecteren van de confidentiële aard van het eindwerk veroorzaakt onherstelbare schade aan de UGent. Ingeval een geschil zou ontstaan in het kader van deze verklaring, zijn de rechtbanken van het arrondissement Gent uitsluitend bevoegd daarvan kennis te nemen. Acknowledgements ACKNOWLEDGEMENTS Toen ik halverwege het eerste semester plots vernam dat ik van thesisonderwerp moest veranderen, was ik toch wel wat in paniek. Al het werk van voordien leek plots voor niets geweest. Achteraf bekeken was dit gevoel onterecht en kijk ik heel tevreden terug naar het afgelopen thesisjaar. Zonder de hulp van tal van mensen zou dit alles nooit mogelijk geweest zijn. Zij verdienen hier daarom een speciaal woordje van dank. Allereerst mijn promotor, Maxime. Bedankt voor al je advies en tips. Je opmerkingen waren steeds terecht en hebben me gestuurd tot een eindresultaat waar ik best wel trots op ben. Wat zou een thesis verder zijn zonder een fantastische begeleider? Bedankt Stijn voor alle uitleg bij de experimenten, het nalezen van mijn teksten en alles wat je het voorbije jaar gedaan hebt om deze thesis te doen slagen! Natuurlijk ook ontzettend bedankt voor de taart en de pittaschotel. Je hebt er wel een mooie databank van je artikels aan overgehouden, niet? Bij deze wil ik je alvast ook veel succes wensen met het schrijven van je doctoraat. Ook een woordje van dank aan Wouter en Freija voor alle hulp wanneer er problemen waren en aan Marjolein voor het gieten van de plaatjes voor mijn culturen. Myriam, bedankt om me kennis te laten maken met de wereld van de elektronenmicroscopie. Ik ben bijzonder blij dat ik deze techniek van jou heb mogen leren. Ook bedankt voor het vele snijwerk en de fixaties die je uitvoerde voor mijn thesis. Geen coupe was je te veel en je stond altijd voor me klaar. De leuke sfeer op de bureau zal ik ook niet snel vergeten. Pam, Hanne en Barbara met jullie was het altijd ontzettend fijn! Bovendien ook bedankt Pam voor de positieve feedback na het lezen van mijn inleiding. Dank gaat ook uit naar Ineke. Het was aangenaam om jou als medestudent in het labo te hebben. Bedankt voor al het overleg, voor het nalezen van mijn Nederlandstalige samenvatting en ook om er voor te zorgen dat er altijd verse BSA-T klaarstond als ik nog maar eens een kleuring moest uitvoeren. Verder wil ik ook nog het labo bedanken om vorig jaar me de unieke kans te geven om mee te gaan naar het platwormencongres te Hasselt. Deze ervaring was de ideale manier om gemotiveerd met het praktische werk van mijn thesis te starten. Een speciaal woordje van dank gaat ook uit naar mijn ouders. Bedankt om mijn studies te financieren en mij de kans te geven om de laatste twee jaar op kot te gaan. Tenslotte wil ik ook nog de persoon bedanken die zoveel voor me betekent, mijn vriendin, Carolien. Na vijf Characterization of the Macrostomum lignano ageing phenotype i Acknowledgements jaar samen aan de universiteit is het einde eindelijk in zicht! Het waren soms hectische dagen en ook al had het je zelf heel erg druk met je eigen thesis, toch kon ik altijd bij je terecht. Je kritische opmerkingen en de vele tips qua lay-out hebben er zeker voor gezorgd dat deze thesis is wat hij geworden is. Zonder jou was het me nooit gelukt! Characterization of the Macrostomum lignano ageing phenotype ii Table of Contents TABLE OF CONTENTS Acknowledgements .................................................................................................................... i Table of Contents .................................................................................................................... iii List of Tables ............................................................................................................................. v List of Figures .......................................................................................................................... vi Part 1: Introduction ................................................................................................................. 1 1.1 Ageing ............................................................................................................................................... 1 1.1.1 Definitions of ageing .............................................................................................................. 1 1.1.2 Causes of ageing ..................................................................................................................... 1 1.1.3 Stem cells and ageing ............................................................................................................. 2 1.2 Ageing model systems ....................................................................................................................... 3 1.2.1 Comparative biology .............................................................................................................. 3 1.2.2 Advantages and disadvantages of current model organisms .................................................. 4 1.2.3 Is there a need for new ageing model systems? ...................................................................... 4 1.3 A flatworm ageing model system ...................................................................................................... 5 1.3.1 Flatworms and stem cells ....................................................................................................... 5 1.3.2 The free-living flatworm, Macrostomum lignano .................................................................. 6 1.3.3 Macrostomum lignano as an ageing model system ................................................................ 7 1.3.4 Ageing in Macrostomum lignano ........................................................................................... 8 1.4 Hypothesis ......................................................................................................................................... 9 Part 2: Objectives ................................................................................................................... 10 Part 3: Material and Methods ............................................................................................... 11 3.1 Overview ......................................................................................................................................... 11 3.2 Ageing cultures ................................................................................................................................ 12 3.3 Light microscopical analysis ........................................................................................................... 12 3.3.1 Principle ................................................................................................................................ 12 3.3.2 Procedure .............................................................................................................................. 12 3.4 Transmission electron microscopy .................................................................................................. 13 3.4.1 Principle ................................................................................................................................ 13 3.4.2 Procedure .............................................................................................................................. 13 3.4.3 Software ................................................................................................................................ 14 3.5 3D-Reconstruction ........................................................................................................................... 14 3.6 Neoblast labelling ...........................................................................................................................
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