Epigenetic Programming by Prenatal Stress in Female Serotonin Transporter Deficient Mice

Epigenetic Programming by Prenatal Stress in Female Serotonin Transporter Deficient Mice

Epigenetic programming by prenatal stress in female serotonin transporter deficient mice Epigenetische Programmierung durch Pränatalstress in weiblichen Serotonintransporter-defizienten Mäusen Doctoral thesis for a doctoral degree at the Graduate School of Life Sciences, Julius-Maximilians-Universität Würzburg, Section Neuroscience Submitted by Karla-Gerlinde Schraut from Timişoara, România Würzburg, 2015 Submitted on: ............................................................................. Members of the Promotionskomitee: Chairperson: Prof. Dr. Thomas Dandekar Primary Supervisor: Prof. Dr. Klaus-Peter Lesch Supervisor (Second): Prof. Dr. Charlotte Förster Supervisor (Third): Dr. Daniel van den Hove Date of Public Defense: ............................................................... Date of Receipt of Certificates: .................................................... Table of Contents Table of Contents ................................................................................................................................ I Abbreviations ..................................................................................................................................... V Summary…………………………………………………………………………………………………..XI Zusammenfassung ........................................................................................................................... XV 1. General introduction ................................................................................................... 1 1.1. Variation in the serotonin transporter gene – from mice and men ................................ 1 1.1.1. Anatomy and function of the serotonergic system .............................................. 1 1.1.2. 5-HT synthesis and metabolism ................................................................................. 3 1.1.3. Dysfunction of the serotonergic system in psychiatric disorders ........................ 3 1.1.4. Genetic variation in the 5-HTT gene ........................................................................ 4 1.1.5. 5-HTT deficient mice ................................................................................................... 6 1.2. Prenatal stress ............................................................................................................................ 9 1.2.1. Early environment might shape adult phenotypes .............................................. 9 1.2.2. What do we know about the effects of prenatal stress? ..................................10 1.2.3. The HPA axis ...............................................................................................................13 1.2.4. Stress hormones during pregnancy .......................................................................16 1.3. Epigenetics ..............................................................................................................................19 1.3.1. DNA methylation and the epigenome .................................................................19 1.3.2. Functional molecular mechanisms of DNA methylation and demethylation .....................................................................................................................................21 1.3.3. DNA methylation patterns across the lifespan ....................................................23 2. Project 1 – The effects of 5-Htt genotype and PS on hippocampal gene expression and DNA methylation ...........................................................................27 2.1. Introduction .............................................................................................................................27 2.2. Methods and Materials .........................................................................................................29 2.2.1. Animals and ethics ...................................................................................................29 I 2.2.2. Behavioral testing .....................................................................................................29 2.2.3. Gene expression analysis .........................................................................................31 2.2.4. DNA methylation analysis ........................................................................................34 2.2.5. Statistics .......................................................................................................................39 2.3. Results .......................................................................................................................................41 2.3.1. Expression of myelin-associated genes.................................................................41 2.3.2. Gene expression of major myelin proteins and transcription factors in hippocampus and amygdala assessed by RT-qPCR .........................................47 2.3.3. DNA methylation .......................................................................................................52 2.4. Discussion .................................................................................................................................69 2.4.1. Effects of 5-Htt genotype and PS on DNA methylation .....................................69 2.4.2. Genes affected both on expression and DNA methylation level....................70 2.4.3. 5-Htt genotype dependent changes in expression of myelin-associated genes and in Mbp methylation induced by PS ..................................................72 2.4.4. On the (missing) relationship of DNA methylation and expression ..................77 2.4.5. Study limitations .........................................................................................................79 2.4.6. Conclusion and outlook ..........................................................................................80 3. Project 2 – Resilience towards PS in 5-Htt deficient female mice .....................83 3.1. Introduction .............................................................................................................................83 3.2. Methods and Materials .........................................................................................................85 3.2.1. Animals and ethics ...................................................................................................85 3.2.2. Behavioral testing .....................................................................................................85 3.2.3. Physiological parameters ........................................................................................87 3.2.4. Processing of brain tissue .........................................................................................88 3.2.5. Gene expression analysis .........................................................................................89 3.2.6. Statistics .......................................................................................................................90 3.3. Results .......................................................................................................................................91 3.3.1. Dam and pups weight .............................................................................................91 3.3.2. Offspring behavior ....................................................................................................91 3.3.3. Physiological parameters ........................................................................................97 II 3.3.4. Transcriptome analysis using mRNA sequencing ................................................98 3.4. Discussion .............................................................................................................................. 157 3.4.1. Behavioral effects of 5-Htt x PS interaction ....................................................... 157 3.4.2. Hippocampal gene expression profiles of 5-Htt+/- mice exposed to PS ..... 159 3.4.3. Altered expression of genes associated with myelination and oligodendrocytes due to 5-Htt x PS interaction ............................................... 163 3.4.4. Manipulation at the 5-Htt locus affected expression of numerous genes on chr 11 ....................................................................................................................... 169 3.4.5. Pathways affected in social, but not in unsocial, 5-Htt+/- mice exposed to PS .................................................................................................................................. 173 3.4.6. Study limitations and methodological issues .................................................... 176 3.4.7. Conclusion and outlook ....................................................................................... 177 4. Appendix .................................................................................................................. 180 4.1. Appendix Tables (see enclosed DVD) ............................................................................. 180 4.2. References ............................................................................................................................ 181 4.3. List of figures ......................................................................................................................... 202 4.4. List of tables .......................................................................................................................... 204 4.5. Curriculum vitae .................................................................................................................

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