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Effects of Gonadotropin-Releasing Hormone Agonist on Brain Development and Aging: Results from Two Animal Models

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Effects of Gonadotropin-Releasing Hormone Agonist on Brain Development and Aging: Results from Two Animal Models Effects of gonadotropin-releasing hormone agonist on brain development and aging: results from two animal models Syed M. Nuruddin Thesis for the Degree of Doctor of Philosophy (PhD) Faculty of Veterinary Medicine and Biosciences Department of Production Animal Clinical Sciences Oslo, 2013 1 CONTENTS Acknowledgment ......................................................................................................................................................... 4 Summary ...................................................................................................................................................................... 6 Sammendrag (Norwegian) .......................................................................................................................................... 9 Abbreviations ............................................................................................................................................................. 12 List of papers .............................................................................................................................................................. 15 Introduction ............................................................................................................................................................... 16 Gonadotropin- releasing hormone and its receptor ................................................................................................ 17 Functional roles of mammalian GnRH and its receptors ........................................................................................ 18 Functions in the reproductive system .................................................................................................................. 18 Function in the central nervous system ............................................................................................................... 20 Functional roles of hippocampus and amygdala in cognition ................................................................................ 22 Hippocampus ...................................................................................................................................................... 22 Amygdala ............................................................................................................................................................ 23 Role of GnRH and its receptor in cognitive function .............................................................................................. 25 GnRH agonist .......................................................................................................................................................... 26 Application of GnRH agonist in adults and their side effects ............................................................................. 26 Usage of GnRHa in pediatric medicine .............................................................................................................. 27 Aim of the thesis ......................................................................................................................................................... 29 Materials and Methods ............................................................................................................................................. 31 Animals and treatment ............................................................................................................................................ 31 Methods ................................................................................................................................................................... 33 Behaviour study - Spatial orientation task .......................................................................................................... 34 Hippocampal gene expression involved in synaptic plasticity and endocrine signaling .................................... 35 Postmortem magnetic resonance image (MRI)) for morphometric analysis of global and regional brain volumes ............................................................................................................................................................... 36 Transcription profiling through microarray in amygdala samples .................................................................... 38 2 mRNA Expression of GnRHI and GnRHR in the hippocampus of Transgenic mice ........................................... 40 Measurement of amyloid-β deposition in cerebral cortex, thalamus and hippocampus using immunohistochemistry ........................................................................................................................................ 40 Statistical analyses .............................................................................................................................................. 42 Results; summery of the papers ............................................................................................................................... 46 Paper I ................................................................................................................................................................ 46 Paper II ............................................................................................................................................................... 47 Paper III .............................................................................................................................................................. 48 Paper IV .............................................................................................................................................................. 49 Discussion ................................................................................................................................................................... 50 Methodological considerations ....................................................................................................................... 51 Animal models ................................................................................................................................................. 51 Spatial orientation test ........................................................................................................................................ 54 Microarray .......................................................................................................................................................... 55 Quantitative real time pcr (qRT-PCR) ................................................................................................................ 57 Magnetic resonance image (MRI) ....................................................................................................................... 57 Immunocytohistochemistry ................................................................................................................................. 59 Experimental findings ............................................................................................................................................. 60 GnRHa treatment impact in spatial orientation as well as hippocampal genes involved in synaptic plasticity and endocrine signaling ...................................................................................................................................... 60 GnRHa treatment effect on amygdalar volume of the brain ............................................................................... 63 Impact of GnRHa treatment on gene expression changes in the amygdala ........................................................ 66 GnRHa treatment effect on gene expression changes and amyloid plaques deposition ..................................... 69 Relevance of findings for human medicine ......................................................................................................... 70 Conclusions ................................................................................................................................................................ 71 Further studies ........................................................................................................................................................... 72 References .................................................................................................................................................................. 74 Paper I-IV 3 Acknowledgment I want to take this opportunity to thank my colleagues, family and friends for their support over the last few years throughout my PhD, and also to thank the Norwegian School of Veterinary Science, Norwegian Research Council, by Health Authorities South East of Norway (2007), AstraZeneca London (2008) and Welcome Trust for funding my PhD project. Firstly I would like to thank Associate Professor Anette Krogenæs, Professor Erik Ropstad, Professor Ira Haraldsen, Professor Neil Evans, Professor Jane Robinson and Professor Mahmood Amiry- Moghaddam for all of their supervision, guidance and encouragement throughout my PhD. It has been a pleasure working together for the past four years. Particularly, I am grateful to Ira and Erik for introducing me to the world of scientific research. Ira and Erik, your ambition and passion for science have been truly inspiring. Your guidance throughout the writing process has proven invaluable and has contributed substantially to the quality of my
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