EURON and THEME joint PhD meeting

Transcription

1 September EURON and THEME joint PhD meeting

2 Euron local coordinators Full partners The Netherlands Maastricht University Dr. Jos Prickaerts Germany RWTH Aachen University Dr. Gary Brook Prof. Dr. Jochen Walter Universität Köln Prof. Dr. Hannsjörg Schröder Saarland University, Homburg Prof. Dr. Tobias Hartmann Belgium Katholieke Universiteit Leuven Dr. Tilmann Achsel Université Libre de Bruxelles Prof. Dr. Roland Pochet Université catholique de Louvain Prof. Dr. Pascal Kienlen-Campard Universite of Liège Prof. Dr. Pierre Leprince Hasselt University Prof. Dr. Jean-Michel Rigo Associated partners University of Leipzig, Germany Prof. Dr. Andreas Reichenbach University of Minho, Portugal Prof. Dr. Joana Palha CPN INSERM University Paris Descartes, France Dr. Laurence Lanfumey Ege University, Izmir, Turkey Prof. Dr. Canan Y. Salam Université Lille 1-Sciences et Technologies Prof. Dr. Michel Salzet Ondokuz Mayis University, Samsun, Turkey Prof. Dr. Suleyman Kaplan

3 EURON and THEME joint PhD meeting, Germany September 22-23, 2011 The local organizing committee: Prof. Dr. Jochen Walter Dr. Andrea Weber Prof. Dr. Volkmar Gieselmann Iris Ullrich Euron: Prof. Dr. Harry Steinbusch Dr. Nicole Senden Peggy Bisschoff Marie-Thérèse Moers

4 Welcome of THEME Dear EURON and THEME graduate students, Dear Colleagues, Welcome to the second annual THEME symposium in Bad Honnef, this year proudly hosted together with EURON, the European Graduate School of Neurosciences. This annual symposium has been designed to give PhD-students the opportunity to present their exciting research results to an international audience as either an oral or poster presentation. The curriculum is complemented by cutting edge talks of senior scientists, who will give insights into the challenges of fundamental research and demonstrate how to address and solve the underlying questions. I am grateful and excited holding this years THEME meeting together with EURON, This is a unique chance for an extensive dialogue between young and senior scientists on an international level. I encourage you to be part of lively discussions in the oral presentation as well as for the poster sessions. Take the chance to discuss your results. The scientific dialogue is an important basis for new and stimulating ideas and this in turn is the basis for a successful symposium. I wish all of us a pleasant and rewarding meeting! Prof. Dr. med. Volkmar Gieselmann 2

5 Information about THEME THEME International Graduate School of Theoretical and Experimental Medicine The Faculty of Medicine in collaboration with the Faculty of Mathematics and Natural Science and the University Hospital of Bonn provide an excellent infrastructure for life science oriented research. The scientific and educational framework of THEME is built by renowned groups covering the main research fields of the Faculty of Medicine focused on: Neuroscience Genetics and Epidemiology of Human Diseases Diseases of the Cardiovascular System Research encompasses a broad range of topics from molecular mechanisms in the development and progression of diseases to prevention, diagnosis and treatment. It also includes patient oriented clinical research. Courses of the study programme of THEME include lectures, seminars and practical courses and are integrated into the research fields mentioned above. THEME communicates relevant theoretical and practical skills needed for a successful thesis and prepares for a career in medical and life science research. THEME does not rely on external funding which specifically supports a graduate school. Rather it provides a framework for Ph.D. students working within peer reviewed projects mostly funded by the German Research Foundation (DFG), the EU of the German Ministry of Science. Therefore it is a permanent institution within the Medical Faculty of the which aims to continuously improve the conditions for a succesful accomplishment of an excellent Ph.D thesis. 3

6 Welcome Address of the Euron Director I would like you to welcome you to our fifteenth EURON PhD days organized by the in Bad Honnef. It is for me a memorable event since it is our second EURON meeting that will be organized in Bad Honnef. The first one was at the ocassion of our masterclass/ workshop on Drugs and the Brain. For our new members from the University of Saarland, Homburg, and the University of Lille, I would like to remind that EURON has started already fifteen years ago with the idea to bring together neuroscientists in the Euregio to generate a platform for scientific and educational exchange for Master as well as PhD students and to stimulate a continuous life long learning programme. We initially have started with an educational program for PhD students and we could integrate all expertise of the scientists of our participating institutes. The educational program and the scientific collaborations attract the attention of potential new Master, PhD students and post-docs. Over the last fifteen years we were able to stimulate our collaboration through funding from the EU by two Marie Curie Early Stage Training Sites under the FP5 and FP6 programmes. Last year we received additional funding from the European Erasmus Life Long learning Program to develop within EURON a two years Research Master program European Master in Neuroscience. This Master will attract Master Students from all over Europe which later can start a PhD program within our network. The original educational idea is still valid and is even stronger these days, since nowadays also the EU and FENS have started within the NENS organization a focus on training for PhD students. Our new intitiatives encompass two new projects for which we will apply in 2012 on joint Master and PhD degrees. In addition the exchange of master and PhD students between the various centres is strongly encouraged and will be discussed further. However, it is not our aim to bring all scientific interests together under one common theme, moreover it is not necessary. Each of the participating institutes have their specific expertise which we will use in our joint applications. Scientifically EURON is moving from systems neuroscience towards more translational neuroscience. During this joint meeting with THEME we will have 29 oral presentations and 40 poster presentations of the PhD students of our Graduate schools. We are pleased to offer jointly this platform to present your data to colleagues within 4

7 the Graduate Schools and more important we hope you will use the opportunity to start your own networking. EURON can provide each of you with a EURON certificate on top of your local degree. This extra certificate will give you a premium on top of your local PhD degree and therefore better possibilities on the later post-doc market. It makes you special and it shows your premium quality. Information on the requirements to obtain this EURON certificate can be found in this booklet and on our website. We encourage all students to go for 3 to 6 months abroad to another lab and add or continue your research project within EURON. We offer opportunities, however it is you to decide to make use of them to your advantage. I wish you two scientific rewarding and also socially exciting days in Bonn / Bad Honnef. Prof. Dr. Harry Steinbusch, Director of EURON 5

8 6

9 EURON a short introduction 7

10 Information from EURON EURON currently consists of 10 full partner universities in Belgium, Germany and the Netherlands and 6 associated university partners in France, Portugal, Turkey and Germany. The research school EURON was officially established and accredited in 2003 and successfully re-accredited in 2010 by the Royal Netherlands Academy of Arts and Sciences (KNAW). Euron has received at July 2010 its official approval (with delight) of this follow up recognition. The Committee was praising the international orientation and the way at which EURON tries to link the research and educational collaboration between the partners. This recognition is again for a six years period. Furthermore EURON has a strong international position due to its recognition by the EU as Marie Curie Early Stage Training Site during the period Our new initiatives are: To develop a joint Research Master program in Neuroscience between the EURON partner universities supported by a Curriculum Development project of the Erasmus Lifelong Learning Program. Estimated start date: September This joint master program will result in a joint degree. For Euron the EMiN program is the best example to share knowledge and expertise between universities and countries. To (re) apply for a FP7 People Marie-Curie Training Network program with our SYNERGY project Interdisciplinary Training Network in New Therapeutic Targets for Neurodegeneration (January 12th, 2012). Innovative treatments can only result from innovative research. Therefore EURON has developed a new initiative in which seven universities and six large companies will work together to provide insight into the neurodegenerative processes and disorders in order to increase the development of new therapeutic strategies to prevent or slow down neurodegenerative processes. The complementary network of industrial and academic partners will combine their industrial or scientific knowledge: expertise and resources with a particular focus on the exchange of molecular, anatomical, electrophysiological techniques in neuroscience. The realizations of joint PhD projects between EURON partner universities. In 2009 EURON formally realized a so-called Rotationsstellen (PhD projects) between Maastricht University and the Klinikum Aachen and these joint PhD projects will now be also established between Maastricht University and Hasselt University called sandwich positions. 8

11 To apply for an Erasmus Mundus joint Doctorates April 2012 with the theme: CNS repair. Coordinators Hasselt and Maastricht University. EURON invests in a scientifically stimulating and inspiring environment for Master, PhD students and Post docs to perform disease oriented research and to be part of our extensive educationa program that reflects the expertise of our universities. EURON aims: To stimulate mobility of students and teachers To share knowledge and expertise To transfer knowledge To aim for uniformity of PhD degrees To agree to the Bologna and Lisbon process The EURON Certificate is the way EURON encourages amongst others the uniformity in PhD degrees. Furthermore it will provide the EURON PhD students an extra recognition in their curriculum and that will be of advantageous for their further career. To obtain this certificate is depending on the following requirements: PhD thesis in English language Obligation to follow courses and to obtain credit points (based on the ects system: in 2012 this will be implemented. 10 ects from the EURON course program according to the entire PhD) One EURON member (not from the home university) should represent EURON on the dissertation review committee and/or at the dissertation defense. Scientific exchange visit(s) to other EURON research groups for a period of at least 3 months, related to the PhD program or other foreign experience. The thesis should consist of at least four papers or of papers with a total Impact Factor of at least 10 as first-author or joint first author. Adjusted weight factors for the various disciplines in neuroscience are considered Admittance to the EURON PhD degree program is limited to individuals holding a MSc, MD or MA degree or equivalent. 9

12 EURON courses and events Autumn November 21, 2011 November 24, 2011 January 26-27, 2012 February 6-9, 2012 April 16-20, 2012 Sept. - Oct Oct. Nov th MHeNS Translational Workshop on Pain School for Mental Health and Neuroscience (MHeNS)/ Maastricht University EURON Course Rodent Neuroanatomy University of Cologne Euron course on Stereology in Neuroscience. Maastricht University / Ondukuz Mayis University, Samsun / Ankara University MHeNS Course From Neuroanatomy to Psychopathology MHeNS / Maastricht University FENS-IBRO/EURON Workshop Drugs and the Brain: an update in Psychopharmacology Maastricht University / University of Minho Sponsored by the ECNP 16th EURON PhD Days Maastricht University (dates need to be defined) Update on Alzheimer Research: Workshop for PhD-students New Orleans, USA, Preceding SFN 2012 EURON / ISAO / AHAF Information and registration courses and events: 10

13 List of EURON partner universities that have organized the EURON PhD Days in the past. 2000: Maastricht University 2001: Katholieke Universiteit Leuven 2002: University of Cologne 2003: Université de Liège 2004: Saarland University, Homburg 2005: Université de Liège 2006: Maastricht University 2007: Université catholique de Louvain 2008: RWTH Aachen 2009: Radboud University Nijmegen 2010: Hasselt University 2011: 11

14 12

15 Programme 13

16 Location: Seminaris Hotel Bad Honnef Wednesday September 21 st 13:00-13:20 Registration 13:20-13:30 Opening and Welcome 13:30-15:30 Session I (Chair: W. Kunz) O. Braganza Illuminating recurrent circuits in the epileptic dentate gyrus: a population Ca 2+ imaging study J. Anschlag Tcfap2c target genes in mouse primordial germ cells J. Lodder Characterisation of two N-acetylaspartylglutamate synthetases R. Hardt Functions of fatty acid 2-hydroxylation in mammals D. Gerlach Intracerebroventricular enzyme infusion corrects central nervous system pathology and dysfunction in mouse model of metachromatic leukodystrophy K. Schulte Simultaneous activation of the presynaptic cannabinoid CB 1 receptor attenuates the function of the presynaptic muscarine M 2 receptor and the δ opioid receptor 15:30-16:00 Coffee/tea 16:00-17:40 Session II (Chair: V. Gieselmann) M. Trautmann Inhibition of WNT signaling impairs growth of synovial sarcoma cells T. Nguyen Presenilins regulate autophagic induction V. Peeva Multiple mitochondrial DNA deletions in human diseases A. Bock The extracellular allosteric area of a seven transmembrane receptor controls intracellular signal trafficking C. Kilgus Local gene targeting and cell positioning using magentic nanoparticles for the generation of biological cardiac pacemakers Final Discussion Walking Dinner/Aperitive 14

17 Thursday September 22 nd 9:00-10:00 Registration and poster set-up 10:00-10:10 Opening and Welcome Prof. Dr. med. Volkmar Gieselmann and Prof. Dr. Jochen Walter 10:10-10:15 Communications Prof. Dr. Harry Steinbusch 10:15-11:00 Key lecture Prof. Dr. Eckhard Mandelkow MPI Hamburg/German Center for Neurodegenerative Diseases Bonn Structural principles of Tau aggregation and Tau-dependent neurodegeneration (Chair: J. Walter) 11:00-12:20 Session I - Neurodegeneration (Chair: P. Kienlen-Campard) R. Gentier Expression of aberrant ubiquitin B (UBB +1 ) in the brainstem of a transgenic mouse model with AD-associated phenotype F. Dennissen Mutant ubiquitin (UBB +1 ) associated with neurodegenerative disorders is hydrolysed by UCH-L3 C. Mencarelli The role of Goodpasture antigen-binding protein (GPBP) in the cellular response against Aβ-induced toxicity S. Tosheva γ-secretase dependent phagocytosis of Amyloid-beta (Aβ) in microglial cells 12:20-13:30 Lunch 13:30-14:15 Key lecture Prof. Dr. Frank Kirchhoff University of Saarland Mechanisms of neuron-glia interaction in vivo what transgenic mouse models tell us (Chair: H. Steinbusch) 15

18 14:15 15:35 Session II Neurodegeneration (Chair: M. De Baets) A. Sierksma Persistent spatial memory improvement after phosphodiesterase type 4d inhibition in the APPswe/PS1dE9 mouse model of Alzheimer disease T. Vanmierlo Brain-derived neurotrophic factor (BDNF), a bridge between depression and Alzheimer s disease D.H. Zeef Memory deficits in transgenice huntington's disease rats R. Nalavade Investigating the role of mirnas in Spinocerebellar Ataxia type 3 (SCA3) 15:35 16:00 Coffee/tea 16:00 17:00 Session III Behaviour (Chair: H. Beck) I. Carvalho Melo Role of Penk gene in stress reactivity I. Rayen Maternal fluoxetine exposure, regardless of prenatal stress, affects physiological systems involved in sexual development of offspring M.E. Siwek The influence of spatial distortion during body perception: an event-related potential study 17:00 18:20 Session IV Neurocommunication (Chair: J-M. Rigo) S. Paßlick Does the proteoglycan NG2 influence neuron-ng2 cell synaptic signaling? M. Vaessen Aberrant modular organization of cerebral functional networks in cognitive impaired children with frontal lobe epilepsy L. Pothmann Effects of antiepileptic drugs on hippocampal inhibitory microcircuits in the epileptic hippocampus C. Albus Altered hippocampal network oscillations in ASA-deficient mice Aperitive 19:00 Walking Dinner + Posters 16

19 Friday September 23 rd :40 Session V Neurocommunication (continued) (Chair: C. Steinhäuser) H. Dannenberg Labelling and optogenetic manipulation of learning-related neuronal assemblies M. Pabst Optogenetic modulation of the septohippocampal pathway 9: Session VI Genetics (Chair: C. Steinhäuser) S. Heilmann Identification of HDAC9 on chromosome 7p21.1 as a new candidate gene for androgenetic alopecia S. Horpaopan Identification of new causative genes in patients with adenomatous polyposis by copy number variation analysis :40 Coffee/tea 10:40-12:20 Session VII Neuroinflammation (Chair: M. Theis) A. Maheshwari Local overexpression of Interleukin -11 in the central nervous system prevents demyelination C.-H. Chang Characterization of Kir channel expression in Schwann cells of the sciatic nerve in a mouse model of metachromatic leukodystrophy S. Smolders Migration of microglia in the embryonic neocortex L. Bodea Investigating the Role of Microglial TREM2 Receptor under Normal and Pathological Conditions E. Vlassaks The effects of asphyctic preconditioning and perinatal asphyxia on inflammation 12:20-13:15 Lunch 17

20 13:15-14:55 Session VIII Neurodevelopment (Chair: V. Gieselmann) L. Mürtz Identification of micrornas regulating differentiation of human neural stem cells B. Roese-Koemer The impact of bifunctional microrna-9/9* on the differentiation of human ES cell derived neural stem cells A. Avila Glycine receptor activation influences cortex development G. Bodea Unraveling the migratory behavior of dopaminergic neuronal subpopulations in the ventral midbrain A. Kabanova Determining the role of Shh signaling in establishing midbrain dopaminergic neuron subclasses :15 Awards / Closing remarks 18

21 Poster presentations 1. Sven Akkerman Methodological considerations on exploration and discrimination measures of object recognition 2. Buket Basmanav Rare variants on the schizophrenia associated 1q21.1 microdeletion region 3. Jessica Becker Genome-wide Meta-analysis on Dyslexia 4. René Besseling Tract specific morphological and parametric reproducibility 5. Eva Bollen TrkB agonist 7,8-dihydroxyflavone improves memory in an object recognition task 6. Verena Borm Biochemical and functional properties of Liprinsalpha 7. Claudia Cornelissen Immunohistochemical characterization of glial cells in human temporal lobe epilepsy 8. Kimberly Cox Neurogenomics in perinatal asphyxia and fetal preconditioning 9. Kristina Dobrindt A human ips cell-based model for autosomal dominant hereditary spastic paraplegia 10. Marianne Eisenhardt Extra-hepatic accumulation of CXCR3(+) NK cells as a novel mechanism of dysregulated NK cell function may contribute to immunopathogenesis of chronic HCV infection 11. Daniela Evers Reprogramming of induced pluripotent stem cells from human cord blood 12. Stephanie Friedrichs Cardiomyocytes obtained from murine induced pluripotent stem cells with long QT syndrome 3 recapitulate typical disease-specific features in vitro 19

22 13. Mary Gazea The role of primary cilia in the development of dopaminergic neurons in the murine ventral midbrain 14. Jose Gerardo Nava Development of an in vitro system for the assessment of axon growth promoting properties of bioengineered scaffolds 15. Andreas Glässner Natural killer cells from HCV-infected patients are highly efficient in inducing apoptosis of activated primary human hepatic stellate cells 16. Alejandro Gomez Repair mechanisms at the neuromuscular junction. The role of Dok7 17. Michaela Granzow Role of the Renin-Angiotensin-System in fibrosis and cirrhosis with portal hypertension in rats 18. Stephanie Griemsmann Morphological and functional analysis of astrocytes in the thalamus 19. Caroline Hammels Susceptibility and resilience in the social defeat model 20. Verena Herl Bidirectional expression of Lck-GCaMP3 and DsRed in NG2 cells as an approach for monitoring glial Ca 2+ - microdomains 21. Katia Herz Monitoring of vascular development in the murine embryonic stem cell system using a flt-1/ egfp construct 22. Dorothee Hodde Development of a Nanofibre-Based Tissue Engineering Strategy to Promote Functional Repair Following Traumatic Nervous Tissue injury 23. Ali Jahanshahi Motor cortex stimulation induced neurogenesis 20

23 24. Anna-Lena Klauke Targeting neuropathic pain by endocannabinoid signaling 25. Sabine Klein HSC directed inhibition of Rho-kinase reduces portal pressure without systemic effects 26. Sylvie van der Kruijs Abnormal functional connectivity between areas involved in emotion and executive control in psychogenic non-epileptic seizures (PNES) 27. Andreas Lindstrot Gene expression profile of laser-microdisected prostate cancer tissues 28. Roopika Menon Determining the Protein Profile of Prostate Cancer samples harboring the ERG rearrangement using MALDI Imaging Mass Spectrometry 29. Kim Neitzert The role of the C-C chemokine CCL17 in Alzheimer s disease 30. Neville-Andrew Niessen Effect of prenatal stress and developmental fluoxetine exposure on hippocampal glucorticoid receptors and coactivator GR 31. Astrid Ooms Function of BAG-3 in Skeletal and Cardiac Muscles 32. Annika Ottersbach Nanomagneto-assisted cell tracking in vitro 33. Geke Overvliet Cortical abnormalities and language impairment in Rolandic epilepsy 34. Lutz Priebe Association between copy number variants in 16p11.2 and major depressive disorder in a German case-control sample 35. Sarah Rieck The application of different magnetic nanoparticles optimizes lentiviral transduction and cell positioning of endothelial cells 21

24 36. Karin Rohleder Activity-dependent expression of presynaptic SV2 proteins in hippocampal synapses 37. Anna Schueth Retrograde tracing of neurons in the rodents lateral bladder wall - An experimental study with fluorescent latex beads: mucosal layers of the bladder vs. small intestinal mucosa 38. Jo Stevens Cloning and production of anti inflammatory antibodies against A-BETA 39. Svenja Ternes Generation of conditional knock out mice for diacylglycerol lipase α and β 40. Sarah Vosen Radially symmetric endothelial cell replacement and lentiviral targeting in vessels by the use of magnetic nanoparticles (MNPs) 22

25 EURON Lecture Frank Kirchhoff ADDRESS Saarland University Department of Molecular Physiology Institute of Physiology Building Homburg Germany frank.kirchhoff@uks.eu Homepages: Phone: +49 (0) Fax: +49 (0) Birthdate: POSITION Professor of Molecular Physiology 1. Education 1986 University of Hannover Diploma (Biochemistry) 1990 University of Heidelberg Dr. rer nat. 2. Positions and employment University of Heidelberg - Institute of Neurobiology Postdoc with Helmut Kettenmann, Heidelberg, Germany Max-Delbrück-Centrum for Molecular Medicine, Cellular Neurosciences, Berlin, Germany Postdoc and project leader with Helmut Kettenmann, Berlin 23

26 Max Planck Institute of Experimental Medicine, Department of Neurogenetics, Göttingen, Germany Research group leader Glial Physiology and Imaging Saarland University, Medical Faculty, Institute of Physiology Full Professor for Molecular Physiology 3. Other experience and professional memberships Editorial board Glia, since 2009 Selection committee Studienstiftung des Deutschen Volkes, and mentor Member German Physiological Society, German Neuroscience Society, German Society of Biochemistry, Society for Neuroscience 4. Honours Fellowship Studienstiftung des Deutschen Volkes Fellowship Boehringer Ingelheim Fonds 5. Fields of Specialization Molecular and cellular mechanisms of neuron-glia interactions, transgenic mouse models, in vivo-imaging 24

27 ABSTRACT Mechanisms of neuron-glia interaction in vivo what transgenic mouse models tell us Our research focuses on the molecular and cellular mechanisms of neuron-glia interaction in the central nervous system. We are pursuing two main research questions: How do glial transmitter receptors sense and modulate synaptic transmission? What is the impact for living organisms? How do glial cells respond to acute injuries within the central nervous system? For functional analysis we generated (and are still continuing to develop) transgenic mouse models with cell-type specific expression of various fluorescent proteins (FPs) and inducible gene deletion. We are applying a combination of biochemical and molecular biological methods together with imaging techniques such as two-photon laser-scanning microscopy (2P-LSM) oder CCD imaging. 25

28 26

29 THEME Lecture Eckhard Mandelkow POSITION Director and Scientific Member, Max-Planck-Society 1.Education Year Institution Degree Field of Study Technical University of Braunschweig Vordiplom Physics Tulane University, New Orleans Physics University of Hamburg Diplom Physics Max-Planck-Institute for Med. Research, Heidelberg Ph.D. Biophysics 2. Positions and Honors Postdoctoral fellow, Brandeis University, Waltham MA, USA Group leader, Max-Planck-Institute for Medical Research, Heidelberg Director, Max-Planck-Unit for Structural Molecular Biology, Hamburg Principal Investigator, DZNE, Bonn (German Center for Neurodegenerative Diseases),. Professor, University of Hamburg Scientific Member, Max-Planck-Society 2010 Metlife Award 2011 Potamkin Award 27

30 3. Research fields Structure and function of the cytoskeleton, with special emphasis on Structure of microtubules, actin filaments, intemediate filaments. Structure of microtubule-associated proteins, including Tau. Structures of protein kinases regulating microtubules. Structures of microtubule motor proteins. Axonal transport mechanisms. Cell models of Tau pathology and neurodegeneration. Screening and development of Tau aggregation inhibitors for therapy. Publications Mandelkow, E., Mandelkow, E.-M., Hotani, H., Hess, B., Müller, S.C. (1989). Spatial patterns from oscillating microtubules. Science 246, Wille,H., Drewes,G., Biernat,J., Mandelkow,E.-M., Mandelkow, E. (1992). Alzheimer-like paired helical filaments and antiparallel dimers formed from microtubule-associated protein tau in vitro. J. Cell Biol. 118, Drewes, G., Ebneth, A., Preuss, U., Mandelkow, E.-M., Mandelkow, E. (1997). MARK - a novel family of protein kinases that phosphorylate microtubuleassociated proteins and trigger microtubule disruption. Cell 89, Kozielski, F., Sack, S., Marx, A., Thormählen, M., Schönbrunn, E., Biou, V., Thompson, A., Mandelkow, E.-M., Mandelkow, E. (1997). The crystal structure of dimeric kinesin and implications for microtubuledependent motility. Cell 91, von Bergen, M., Friedhoff, P., Biernat, J., Heberle, J., Mandelkow, E.-M., Mandelkow, E. (2000). Assembly of tau protein into Alzheimer paired helical filaments depends on a local sequence motif (306-VQIVYK-311) forming beta structure. Proc. Natl. Acad. Sci. USA 97, Bulic, B., Pickhardt, M., Schmidt, B., Mandelkow, E.-M., Waldmann, H., Mandelkow, E. (2009). Development of Tau aggregation inhibitors for Alzheimer disease. Angew. Chemie Int. Ed. 48,

31 ABSTRACT Structural principles of Tau aggregation and Taudependent neurodegeneration Eckhard Mandelkow DZNE, German Center for Neurodegenerative Diseases, c/o CAESAR, Bonn, Germany Tau is an unusual protein from several points of view: (1) Neurodegenerative diseases: Changes in Tau are early markers of AD and other "tauopathies" (e.g. FTD, Pick disease, PSP etc), (2) Neuronal cell biology: Tau is a brain-specific microtubule-associated protein (MAP), mostly confined to neuronal axons and implicated in neuronal differentiation, (3) Protein structure: Tau is the prototype of a "natively unfolded" or "intrinsically unstructured" protein, which does not require a defined structure for biological activity (in contrast to most "textbook" proteins with defined secondary and tertiary structure). We are interested in defining the interactions of Tau and the structural basis of its abnormal behavior in neurodegeneration. Tau is best known as an axonal protein that serves to stabilize microtubules, the tracks for long-haul traffic of vesicles and organelles in axons. As such, Tau interacts with tubulin (the building blocks of microtubules), motor proteins (kinesin, dynein), and several protein kinases and phosphatases that regulate these interactions. Malfunction of Tau (e.g. after hyperphosphorylation) can affect the stability of microtubules, interfere with motor-driven transport, and promote the pathological aggregation of Tau after detachment from microtubules. As a natively unfolded protein, Tau is not suitable for high resolution crystallography, however, the structure can be approached by spectroscopies (CD, fluorescence, FTIR, FRET) and NMR (solution and solidstate, collaboration with C. Griesinger, M. Zweckstetter, MPI Göttingen). The spectroscopic studies have revealed that Tau has "hotspots" for aggregation due to their tendency to form β-structure, and that Tau in solution has a "paperclip" folding where the N- and C-termini interact with the repeat domain. Both properties can be modified by phosphorylation at several sites. Knowledge of the β-forming hotspots enables one to design Tau molecules which favor or inhibit aggregation (pro- and anti-aggregant Tau). These variants now form the basis for designing cell- and animal models of tauopathy, and for screening inhibitors of Tau aggregation and other therapeutic agents (collab. B. Bulic, CAESAR). - Supported by MPG, DFG, VW Fnd, BMBF (KNDD), Metlife Fnd. 29

32 30

33 Abstracts 31

34 32

35 Euron PhD meeting 2010 Methodological considerations on exploration and discrimination measures of object recognition S. Akkerman 1,*, A. Blokland 2, O. Reneerkens 1, N.P. van Goethem 1, E. Bollen 1, H.J.M. Gijselaers 1, C.K.J. Lieben 1, H.W.M. Steinbusch 1, J. Prickaerts 1 1 Faculty of Health, Medicine and Life Sciences, Department of Neuroscience, School for Mental Health and Neuroscience, European Graduate School of Neuroscience (EURON), Maastricht University, The Netherlands; 2 Faculty of Psychology and Neuroscience, Department of Neuropsychology and Psychopharmacology, European Graduate School of Neuroscience (EURON), Maastricht University,The Netherlands. The object recognition task (ORT) is a frequently used tool in neurobiological research. In a sample trial, objects are presented to the animal. After a delay interval, one of the sample objects is replaced by a novel object in the test trial. Rats that remember the sample object(s) will spend more time exploring the novel object, compared to the sample object(s), the exploration difference is regarded to be a measure of memory. In this study, 28 ORT experiments were pooled, containing 731 male Wistar rats. We investigated the relationship between 3 commonly used measures of discrimination and how they were related to levels of exploration in the sample and test trial. In this context, the effects of training, trial duration, delay interval and the amnesic drugs MK-801 (0.125 mg/kg) and scopolamine (0.1 mg/kg) on exploration and discrimination measures were investigated. Finally, we addressed object bias and relativity of object novelty as possible factors interfering with exploration and discrimination performance. Our analysis showed that the ORT is sensitive to potential biases like stress and side0-effects of drugs. There was no indication of a relationship between the level of exploration in the sample trial and discrimination performance. On the other hand, exploration in the test trial was positively related to the absolute discrimination measure but not to the relative discrimination measures, making them more resistant to exploration biases. Interaction with objects in prior sessions (training) decreased discrimination during subsequent 24 h retention interval testing. Thus, discrimination appears to reflect a lesser degree of familiarity of the novel object relative to the more familiar sample object due to a more recent encounter with the latter, rather than true novelty per se. Taken together, our findings suggest the consideration of pre-experimental exposure to objects, habituation to treatment procedures, balancing of object presentation and the use of relative discrimination measures when using the ORT. 33

36 Altered hippocampal network oscillations in ASAdeficient mice Christina Albus 1, Matthias Eckhardt 2, Volkmar Gieselmann 2, Heinz Beck 1, Thoralf Opitz 1 1 Laboratory for Experimental Epileptology, Department of Epileptology, Bonn, Germany; 2 Institute of Biochemistry and Molecular Biology, Bonn, Germany. Arylsulfatase A (ASA) is a lysosomal enzyme catalyzing the degradation of sulfatides. Mice deficient in ASA accumulate sulfatides in glial cells, microglia, and neurons and show neuromotor deficits as well as mild behavioral disturbances. In addition, invasive EEG recordings have revealed a marked cortical hyperexcitability, with episodes of spontaneous epileptiform activity. The phenotypic abnormalities have so far been mainly ascribed to the progressive demyelination and axonal damage, but if sulfatide accumulation exerts direct effects on neuronal excitability is so far unknown. To address this question, we first examined excitability on the network level in the hippocampal slice preparation. In the hippocampal CA1 and CA3 subfields, spontaneous slow episodes of population activity were observed under conditions of slightly increased excitability, termed sharp waves (SPW), but their incidence was not different in ASA-deficient mice compared to littermate controls. However, we observed a significantly higher fraction of SPWs with superimposed high frequency oscillations in the CA3 subfield of ASA-deficient mice (termed sharpwave ripples, SWRs). To address the cellular basis for increased SWRs, we have used intracellular recordings to examine the functional properties of pyramidal neurons in both the CA1 and CA3 subfields. We could not identify any systematic changes in passive or active membrane properties in hippocampal principal cells when comparing ASA-deficient mice to littermate control animals. These results suggest that ASA deficiency causes a selectively increased propensity to generate high frequency network activity within the subfield. Given the lack of functional changes in principal neurons, we propose that changes in inhbitory micronetworks may mediate the increase in high-frequency synchronized activity in ASA-deficient mice. Supported by the BMBF (Project LEUKONET) 34

37 Euron PhD meeting 2010 Tcfap2c target genes in mouse primordial germ cells Jana Anschlag 1, Dawid Eckert 1, Hubert Schorle 1 1 Department of Developmental Pathology, Institute of Pathology, Bonn University Medical School, Sigmund-Freud-Str. 25, Bonn, Germany. The transcription factor Tcfap2c (AP-2γ) is a member of the activating protein 2 (AP-2) family of transcription factors and consists of a transactivation domain at the amino terminus and a helix-span-helix motif at the carboxyl terminus. The central basic region is responsible for dimerization and together with the helixspan-helix motif for DNA binding. It is known that AP-2 proteins bind to G/C rich regions like the palindromic sequences 5 -GCCN 3/4 GGC-3 or 5 -GCCN 3/4 GGG-3. During mouse embryogenesis Tcfap2c is expressed in primordial germ cells (PGCs) from embryonic day (E) 7.25 until E Lack of Tcfap2c leads to sterile animals in which PGCs are specified but lost around E 8.0. In these PGCs the expression of germ cell markers is down-regulated and genes indicating somatic differentiation are up-regulated. Tcfap2c has been demonstrated to be upregulated in carcinoma in situ (CIS/IGCNU) and seminoma in humans, raising the critical question regarding the biological function and potential target genes of Tcfap2c in germ cells and testicular germ cells tumors. To investigate the function of transcription factor Tcfap2c in the transcriptional network of PGCs we used an in vitro differentiation protocol to generate PGCs. Mouse embryonic stem (ES) cells harbouring a Stella-GFP transgene were differentiated by embryoid body (EB) aggregation. Upon differentiation in EB culture, PGCs were identified and sorted by Stella-GFP signal a germ cell specific marker. Using these in vitro generated PGCs we performed a ChIP-on-chip analysis to search for promoters recognized by Tcfap2c. Preliminary analysis suggests that Tcfap2c regulates and probably reinforces the genetic and epigenetic network that is utilized to setup PGC identity. This experiment will help to further understanding of Tcfap2c in germ cell biology as well as in germ cell tumors. 35

38 Migratory interneurons express functional glycine receptors during early development of the cerebral cortex Ariel Avila 1,2, Pía Vidal 1, Laurent Nguyen 2 and Jean-Michel Rigo 1 1 BIOMED Research Institute, Hasselt University, Agoralaan C, Diepenbeek; 2 Developmental Neurobiology Unit, Centre for Cellular and Molecular Neurobiology, University of Liege, C.H.U. Sart Tilman, Liège, Belgium. The strychnine-sensitive glycine receptor (GlyR) is a member of the ligand-gated ion channel superfamily. In the adult, the GlyR is known to mediate fast inhibitory neurotransmission in the spinal cord and in the brainstem. The GlyR has also been described in the embryonic cortex from embryonic day 19 (E19) where it could participate in developmental processes, but its presence at earlier stages has not been documented. Since other neurotransmitter systems, i.e. GABA and its receptors, are known to be present during corticogenesis, we wondered if this could also be the case for glycine and its GlyR. In this study, we analyze the presence and physiological relevance of GlyR in the early development of the cortex using in vitro and ex vivo cultures of slices, patch-clamp, two photon microscopy for time lapse and for calcium imaging, immunocytochemistry and western-blot. Electrophysiological experiments confirmed the presence of GlyR mediated currents in migrating interneurons during early stages of development (E13-E15). Using in vitro labeling of interneurons we have described the pharmacological properties of glycinergic currents present in interneurons born at E13. The concentration-response curve showed an EC50 of 69 ± 12 micro M for glycine. All these currents were fully blocked by strychnine with an IC50 of 0.10 ± 0.02 micro M. Picrotoxinin and picrotin also blocked these currents, but with different potency, remaining 20 % of the current when 10 micro M of picrotin was used. Similar glycinergic currents were also observed in ex vivo preparations from Dlx5/6-Cre EGFP transgenic animals, where it was clear that GlyR expressing cells are a subpopulation of migratory interneurons. Consequently, immunostainings directed against the alpha 2 subunit of GlyR showed that 29 ± 2 % of cortical migrating interneurons, which were mainly born in the medial ganglionic eminence (MGE) at E13, express GlyR. All this evidences shows that GlyR appears earlier than ever described during cortex development and it is composed, mainly, by alpha 2 homomeric channels. It also shows that GlyR is not homogenously expressed and it is only present in a subpopulation of migrating interneurons born at a defined space-temporal window during brain development. In search for the physiological function of GlyR, two photon time lapse analysis for cell migration and calcium imaging was performed on ex vivo slices. All these studies 36

39 were complemented by gain and loss of function experiments. As it has been previously described for the GABA system, we show here that GlyR play a role acting on the migratory behavior of interneurons and its effects are linked to modulation of calcium dynamic and the activity of calcium downstream targets. More in detail molecular mechanisms were analyzed by western-blot. 37

40 Tract specific morphological and parametric reproducibility RMH Besseling 1,2,3, JFA Jansen 1,2, GM Overvliet 1,2,3, MJ Vaessen 1,2,3, PAM Hofman 1,2,3, AP Aldenkamp 1,2,3, WH Backes 1,2 1 Department of Radiology; 2 School for Mental Health and Neuroscience, Maastricht University Medical Center (MUMC+), Maastricht, the Netherlands; 3 Epilepsy Center Kempenhaeghe, Heeze, the Netherlands. Rationale Tractography based tract segmentations were used to study reproducibility characteristics of tract volume as well as diffusion tensor metrics at the tract level. In addition, the relatively new diffusion contrast tract density imaging (TDI) was studied at the tract specific level. Furthermore, the reproducibility of the shape of the tracts (tract morphology) was assessed, which included the reproducibility of complete tract segmentations (extended) compared to tract segmentation excluding the directly subcortical projections (proximal). Methods Diffusion weighted imaging (DWI) was performed twice in 9 healthy subjects (28±6 y) on an 3T Philips Achieva scanner using: 128 gradient directions, voxel size of 2x2x2 mm 3, and b=1200 s/mm 2. Standard space seed and inclusion ROIs were mapped non-linearly to native DWI space, and subsequently constrained spherical deconvolution (CSD) probabilistic tractography was performed. The selected tracts were the genu of the corpus callosum, the cingulum, the pyramidal tract, the optic radiation, and the arcuate fasciculus. Tractograms were converted to tract density maps and thresholded to create tract segmentations that were used as ROIs to investigate tract fractional anisotropy (FA), apparent diffusion coefficient (ADC), volume and TDI. In addition, tract segmentations were mapped back to standard space to calculate between-session morphological overlap using the Dice similarity coefficient (DSC). Reproducibility of FA, ADC, volume and TDI was quantified using the coefficient of variation (COV) and the interclass correlation coefficient (ICC). Results For all metrics, reproducibility differed strongly between tracts and between proximal and extended tract segmentations. For FA, the proximal reproducibility values were: COV= %, ICC= For ADC, comparable values were found (COV= %, ICC= ). For volume, proximal COV=6.1-22%, proximal ICC= (0.029 for the arcuate fasciculus). For TDI, proximal 38

41 COV=7.7-27% and not in all tracts accurate ICC values could be estimated. Morphological overlap as expressed by DSC ranged from 0.70 (arcuate fasciculus, extended) to 0.92 (corpus callosum, proximal). For a data subset of 66 gradient directions, DSC values were reduced for each tract with approximately 5%. Conclusion Estimates of tract FA and ADC are reliable, based on favorable COV and ICC values. Tract volume has worse COV values but maintains high ICC values. Tract TDI is the least reliable contrast, due to high COV combined with low ICC. Furthermore, our approach results in very high between-session morphological correspondence (up to DSC=0.92). Reliability is reduced in extended tract segmentations compared to the proximal segmentations. Since both types of segmentations differ primarily in their extent of subcortical projections, a reduced sensitivity in detection of subcortical changes in tract morphology can be expected. 39

42 Conformational strain may underlie biased agonism of dualsteric ligands at the M 2 receptor Bock, A. 1, Müller, A. 2, Holzgrabe, U. 3, De Amici, M. 4, Kostenis, E. 2, Mohr, K. 1 1 Pharmacology and Toxicology Section, Institute of Pharmacy, ; 2 Molecular, Cellular, and Pharmacobiology Section, Institute of Pharmaceutical Biology, ; 3 Institute of Pharmaceutical Chemistry, University of Würzburg; 4 Dipartimento di Scienze Farmaceutiche Pietro Pratesi, Università degli Studi di Milano, Milano, Italy. G protein-coupled receptors (GPCRs) are seven transmembrane (7TM)-spanning proteins representing the largest and most ubiquitously expressed type of cell surface receptors. Many 7TMRs contain at least one allosteric binding site which is topographically distinct from the orthosteric site recognized by the respective endogenous messenger compound. The muscarinic M 2 acetylcholine receptor is an excellent model to study allosteric/orthosteric interactions as the core region of the allosteric binding site is well characterized. Recently, bisquaternary allosteric/orthosteric hybrid compounds were designed consisting of an allosteric inverse agonist fragment linked via an aliphatic hexamethylene middle chain with an orthosteric high affinity agonist. In exclusively activating the Gi pathway these dualsteric compounds showed biased agonism at the M 2 receptor [1]. However, hybrids potency was considerably lower than that of the orthosteric building block alone, suggesting a suboptimal fit of the hybrids building blocks to their corresponding binding sites. Here we show that middle-chain elongation to octamethylene considerably increases potency for M 2 receptor-mediated G protein activation as measured by [ 35 S]GTPγS binding in membranes of hm 2 -CHO cells. In order to check whether middle chain length affects biased signaling we carried out real-time measurements of dynamic mass redistribution in hm 2 -CHO cells using the EPIC system (Corning, New York). Our findings show that the elongated hybrids regain modest ability for G s activation. We conclude that a spatial misfit between a dualsteric ligand and its corresponding orthosteric/allosteric receptor sites might impose a conformational strain on the receptor protein that underlies biased agonism of hexamethylene-type dualsteric compounds. [1] Antony J et al. (2009). Dualsteric GPCR targeting: a novel route to binding and signaling pathway selectivity. FASEB J 23: Support by the DFG is gratefully acknowledged (HO 1368/12-1, MO 821/2-1). This abstract is published in Naunyn Schmiedebergs Arch Pharmacol. 2011, March (383). 40

43 Unraveling the migratory behavior of dopaminergic neuronal subpopulations in the murine ventral midbrain Gabriela O. Bodea and Sandra Blaess Institute of Reconstructive Neurobiology, Neurodevelopmental Genetics Group,, Sigmund-Freud-Str. 25, D Bonn. Midbrain dopaminergic (MbDA) neurons located in the ventral tegmental area (VTA) and the substantia nigra (SN) are involved in many brain functions including reward associated behavior, modulation of emotions and motor control. MbDA neurons are derived from a Sonic Hedgehog (Shh)-expressing precursor domain in the ventral-medial mesencephalon. During development, they have to migrate from this precursor domain to form the laterally positioned SN and the medially located VTA. However, it is not known which migration patterns result in the formation of the distinct MbDA nuclei and which molecular mechanisms direct the migration of MbDA neurons. Using genetic inducible fate mapping to heritably mark Shh-expressing MbDA precursors and their descendants with YFP fluorescent reporter protein, we found that the ventral MbDA precursor domain can be roughly subdivided into a SN and VTA precursor domain. By following the fate of MbDA neurons derived from either the SN or VTA precursor domain during their migration phase (embryonic day ), we are investigating whether the different MbDA subpopulations undergo distinct migration patterns to reach their final position in the SN or VTA. Our analysis of the position and morphology of fate-mapped MbDA neurons at different developmental time points indicates that SN neurons migrate radially while leaving their precursor domain and then switch to tangential migration to reach their final lateral position. In contrast, VTA neurons appear to migrate only radially. To monitor MbDA neuronal migration directly, we established an organotypic slice culture system of the embryonic midbrain, in which we can track the migratory behavior of fluorescently labeled, fate-mapped MbDA neuron subsets using time-lapse imaging. Observation of SN and VTA MbDA neurons in these slice cultures confirms their different migratory behavior. We are now investigating which molecular mechanism regulate radial versus tangential migration of MbDA neurons. 41

44 Investigating the Role of Microglial TREM2 Receptor under Normal and Pathological Conditions Bodea Liviu-Gabriel 1, Linnartz Bettina 1, Colonna Marco 2, Neumann Harald 1 1 Institute of Reconstructive Neurobiology, University Bonn, Bonn, Germany; 2 Washington University School of Medicine, Department of Pathology & Immunology, St. Louis, MO 63110, USA. The triggering receptor expressed on myeloid cells 2 (TREM2) is presented on the cell surface of microglia, the resident immune cell of the central nervous system (CNS). The signaling cascade of TREM2 relies on the DAP12 adaptor molecule and especially on its immunoreceptor tyrosine-based activation motif (ITAM). TREM2/ DAP12 was shown to be involved in the non-inflammatory clearance of apoptotic neuronal material under normal conditions. In humans, a disease caused by loss of function mutation of either TREM2 or DAP12 was identified, the Nasu-Hakola disease, which is characterized by neuroinflammation in the CNS. The present study provides insights on TREM2 function and the mechanism of Nasu-Hakola disease by using a TREM2 knock-out (KO) mouse model. The nigrostriatal system of aged TREM2 KO mice shows signs of mild neurodegeneration. The data also show a slightly increase in microglial Iba1- immunoreactivity in different brain regions of the aged TREM2 KO mice (ventral midbrain, hypothalamus, cortex) compared with the WT animals. However, TNFα, inos and IL-1β cytokine gene transcript levels were unaffected. Preliminary data show that the time frame of neurodegeneration was successfully shortened in TREM2 KO mice by using a protocol based on systemic challenges of mice with inflammatory stimuli,. Thus, injecting lipopolysaccharides (LPS) intraperitoneally on four consecutive days in mice has lead to dopaminergic degeneration but not in the vehicle treated controls. Data show that TREM2 KO mice are more susceptible to degeneration compared to WT controls, thus TREM2 presents a neuroprotective role within the CNS. 42

45 BDNF signaling in hippocampal learning and memory: The effects of 7,8-dihydroxyflavone on object memory E. Bollen 1, J. De Vry 1, T. Vanmierlo 1, H.M.W. Steinbusch 1, J. Prickaerts 1 1 Dept of Psychiatry and Neuropsychology, Maastricht University, The Netherlands. Brain derived neurotrophic factor (BDNF) has emerged as an important regulator of synaptic plasticity in the central nervous system. Binding of BDNF to its main receptor TrkB activates several intracellular signaling cascades, including the PLCγ-Ca 2+ pathway, the Ras-mitogen-activated protein kinase (MAPK) pathway and the phosphatidylinositol 3-kinase (PI3K)-Akt pathways, all of which have been implicated in neuronal growth and/or plasticity. In hippocampal long-term potentiation (LTP), one of the most commonly studied forms of plasticity which is generally considered as the cellular correlate of memory formation, BDNF has been attributed a critical role. In addition, in previous studies we found that the camp- and cgmp-mediated intracellular signaling cascades are exerting their specific effects on object memory consolidation via BDNF. Therefore, we studied the role of TrkB signaling pathways in synaptic plasticity and memory formation by injecting rats with 7,8-dihydroxyflavone (7,8-DHF; 0.3, 1 and 3 mg/kg), a recently identified TrkB agonist during different consolidation phases after learning in an object recognition paradigm. Our results show that 7,8-DHF improves memory when injected both during the early and the late phase of object memory consolidation at 1 and 3 mg/kg. The lowest dose (0.3 mg/kg) resulted in a significant memory improvement only when injected at the late consolidation phase, which may suggest a stronger involvement of BDNF signaling in late memory consolidation. Taken together, these results show the potential of 7,8-DHF as a cognition enhancer. 43

46 Illuminating recurrent circuits in the epileptic dentate gyrus: a population Ca 2+ imaging study Oliver Braganza, Tony Kelly, Heinz Beck Department of Epileptology and Life&Brain Center, University Bonn, Sigmund-Freud-Str. 25, Bonn, Germany. Neural activity is defined by the anatomical and functional connectivity of the underlying neuronal networks. In epilepsy the neural network displays a tendency to spontaneous hypersynchronization, a phenomenon which may well be brought about by changes in the underlying network topology. A hallmark symptom of patients with temporal lobe epilepsy, as well as animal models thereof, is mossy fiber sprouting which is supposed to lead to profound changes in the network structure of the dentate gyrus. Physiologically there is a clear anatomical segregation between the granule cell input zone and their projections, the mossy fibers, reflected in the observation that there is very little interconnectivity between granule cells. In fact, the absence of such recurrent connections within the dentate gyrus granule cell layer is one of its hallmark features. In epilepsy, the anatomical segregation of input and output zone is severely impaired, as mossy fibers grow into the dendritic zone and occasional granule cells with basal dendrites, located in the output zone appear. We suggest that granule cells with basal dendrites exhibit a particularly high interconnectivity within the dentate gyrus, and thus may function as so-called network hubs. Modeling studies have shown that the presence of hub cells with extremely high connectivity alter the dynamics of networks profoundly, and can create hyperexcitability. In order to address questions of network topology it is necessary to simultaneously sample the activity of a large population of neurons with cellular resolution. We describe a multibeam multiphoton Ca 2+ imaging approach that permits fast imaging from large numbers of neurons in adult brain slices. We have used this approach to begin to address changes in network connectivity in the epileptic dentate gyrus. 44

47 Characterization of Kir channel expression in Schwann cells of the sciatic nerve in a mouse model of metachromatic leukodystrophy Cin-He Chang 1, Lihua Wang-Eckhardt 2, Matthias Eckhardt 2, Gerald Seifert 1, Volkmar Gieselmann 2, Christian Steinhäuser 1 1 Institute of Cellular Neurosciences and 2 Institute of Biochemistry and Molecular Biology, Medical Faculty,, Germany Metachromatic leukodystrophy (MLD) is a lysosomal storage disorder caused by deficiency of arylsulfatase A (ASA) and sulfatide storage. ASA deficient mice are an animal model for MLD, which, however, show a relatively mild phenotype. An improved mouse model of MLD, showing increased sulfatide synthesis, has been generated that displays increased sulfatide storage in neural cells of the brain and peripheral nervous system. In the peripheral nerves of these mice, hypo- and demyelination was observed, leading to a slowed propagation of action potentials (Ramakrishnan et al., 2007). Analysis of transcript and protein expression revealed an upregulation of the inward rectifying K + channel subunit Kir4.1 which in the nervous system is expressed solely by glial cells. To investigate whether these changes are accompanied by alterations in Kir4.1 channel function, we applied the patch clamp-technique to Schwann cells freshly isolated from sciatic nerves of ASA-/- mice, ASA-/- mice overexpressing the sulfatide synthesizing cerebroside sulfotransferase in Schwann cells (TG/ASA-/-), and wildtype littermates (WT) (postnatal day ). Schwann cells from WT and ASA-deficient mice displayed depolarized membrane potentials of about -45 mv and a high input resistance (about 400 ΜΩ). These findings did not imply significant expression of functional Kir channels. In line with this assumption, neither WT nor ASA-deficient mice displayed Ba2+ (100 µm)- sensitive inward currents at negative membrane potentials. In conclusion, data obtained so far suggest that in these MLD models, upregulation of Kir4.1 mrna and protein in peripheral nerves is not accompanied by increased expression of functional channels. In the second part of my Ph.D. thesis I m investigating the functional impact of inducible astrocyte-specific ablation of GABA B receptors. Analyses in WT mice revealed that the GABA B R1 subunit is expressed in almost all astrocytes of the hippocampus, whereas pyramidal neurons of the same brain region express both GABA B R1 and GABA B R2 subunits. Recent studies demonstrated that GABA B receptor activation in astrocytes leads to increase in [Ca 2+ ]i, release of gliotransmitters and modulation of synaptic signalling, although the underlying 45

48 mechanisms are not understood. Analysis of GABA B R1 knockout mice will help to better understand how Ca 2+ signaling in astrocytes influences information processing in the brain. 46

49 Neurogenomics in perinatal asphyxia and fetal preconditioning KEM Cox, E Strackx, E Vlassaks, M Sparnaaij, L Zimmerman, JS Vles, AW Gavilanes. Dept of Psychiatry and Neuropsychology, Maastricht University, The Netherlands. Background Part of perinatal hypoxic-ischemic brain damage can be attenuated when it is preceded by fetal preconditioning. The mechanism behind this endogenous neuroprotective phenomenon has not been deciphered yet. It is suggested that genomic reprogramming could be the answer to this question. We performed 50 micro array experiments to evaluate changes in cerebral gene expression after fetal preconditioning and perinatal asphyxia at multiple time points. In this abstract we describe some preliminary results. Objective We aim to analyze the whole genome gene expression at multiple time points in pre- and neonatal pups after fetal preconditioning (FA) and perinatal asphyxia (PA). This can give us insight into the mechanisms of endogenous neuroprotection after FA and the deleterious effects of PA. Unraveling these mechanisms is an important step towards possible new clinical strategies for asphyctic neonates. Design/Methods FA was induced on E17 by clamping the uterine circulation of the maternal rat for 30 minutes. On P0 pups PA was induced by placing the uterine horns, including the fetuses, in a water bath for 19 minutes. Control (CCD) and FA pups were delivered by Caesarean section. These procedures generated four experimental groups: CCD, FA, PA, and FA+PA. Five male pups per group were sacrificed at the following time points: 96h after FA, 6h and 96h after birth. RNA was isolated from the left hemispheres and 50 micro array experiments were performed on the Affymetrix platform. Up- or down-regulation of gene-products was studied via Gene-Ontology terms with a computational method named Gene Set Enrichment Analysis (GSEA). Results With GSEA we found a considerable number of GO-terms that were significantly up- or down-regulated compared to CCD animals (see figure 1). Interestingly in the protected phenotype (FA+PA) no GO-terms were significantly different from the control phenotype 6 hours after birth. Four days after perinatal asphyxia we 47

50 see that numerous GO terms are down-regulated in the PA group. These data will be further analyzed and visualized in pathways; results are expected early September Conclusions These data in rodent fetuses and neonates are the first to demonstrate global gene expression profiles in the brain after fetal preconditioning and perinatal asphyxia. From previous behavioral studies we know that adult FA+PA animals perform similar to adult CCD animals despite the fact that the FA+PA group experienced a perinatal asphyctic insult. Interestingly here we also found no difference between these groups at 6hours after birth. 48