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Long-Term Effects of Immunostimulation on Synaptic Plasticity and Neurodegeneration

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Long-Term Effects of Immunostimulation on Synaptic Plasticity and Neurodegeneration Long-term effects of immunostimulation on synaptic plasticity and neurodegeneration Von der Fakultät für Lebenswissenschaften der Technischen Universität Carolo-Wilhelmina zu Braunschweig zur Erlangung des Grades einer Doktorin der Naturwissenschaften (Dr. rer. nat.) genehmigte D i s s e r t a t i o n von Marianna Martha Simona Beyer aus Braunschweig 1. Referent Professor Dr. Martin Korte 2. Referent Professor Dr. Reinhard Köster 3. Referent Professor Dr. Michael Heneka eingereicht am: 30.11.2015 mündliche Prüfung (Disputation) am: 19.04.2016 Druckjahr 2016 Vorveröffentlichungen der Dissertation Teilergebnisse aus dieser Arbeit wurden mit Genehmigung der Fakultät für Lebenswissenschaften, vertreten durch den Mentor der Arbeit, in den folgenden Beiträgen vorab veröffentlicht: Publikationen Parlog, A., Harsan, L.A., Zagrebelsky, M., Weller, M.*, von Elverfeldt, D., Mawrin, C., Korte, M., Dunay, I.R.: Chronic murine toxoplasmosis is defined by subtle changes in neuronal connectivity, Disease Models & Mechanisms 2014 Apr; 7(4): 459–469. Tagungsbeiträge Weller, M.*, Heneka, M.T., Korte, M.: Long-term Effects of Immunostimulation on Neuronal Morphology, First N-RENNT Symposium on Neuroinfectiology, Hannover (2014) Weller, M.*, Heneka, M.T., Korte, M.: Long-term effects of an immunostimulation on neuronal function and morphology, 9th FENS Forum of Neuroscience, Mailand (2014) Young investigator oral: Beyer, M., Heneka, M.T., Korte, M.: Long-term influences of an immune stimulation on neuronal structure and plasticity, 11th Göttingen Meeting of the German Neuroscience Society, Göttingen (2015) Beyer, M., Heneka, M.T., Korte, M.: Long-term influences of an immune stimulation on neuronal structure and plasticity, Second N-RENNT Symposium on Neuroinfectiology, Hannover (2015) Best Poster Prize: Beyer, M., Heneka, M.T., Korte, M.: Enduring changes in neuronal structure and function upon acute and systemic administration of LPS, 4th Venusberg Meeting on Neuroinflammation, Bonn (2015) Posterbeiträge Weller, M.* Dunay, R.I., Zagrebelsky, M., Korte, M.: Changes in neuronal architecture upon Toxoplasma gondii infection in mice, 4th Public Retreat of the HZI Graduate School, Hahnenklee (2013) Weller, M.*, Heneka, M.T., Korte, M.: Long-term Effects of Immunostimulation on Neuronal Morphology, 6th International phD Symposium of the HZI Graduate School, Braunschweig, (2013) Beyer, M., Heneka, M.T., Korte, M.: Long-term Effects of an Immune Stimulation on Synaptic Plasticity, 7th International phD Symposium, Braunschweig, (2014) Beyer, M., Heneka, M.T., Korte, M.: Enduring changes in neuronal structure and function upon acute and systemic administration of LPS, 6th Public Retreat of the HZI Graduate School, Braunschweig (2015) * Marianna Beyer, geb. Weller Nothing in this world that´s worth having comes easy. Dr. Robert Kelso Für Martin, Martin, Margit, Moritz und Maximilian Table of Contents ZUSAMMENFASSUNG ..................................................................................................... 1 ABSTRACT ....................................................................................................................... 2 1. INTRODUCTION ........................................................................................................... 3 1.1. Innate immunity ....................................................................................................... 3 1.2. Lipopolysaccharide and peptidoglycan-mediated innate immune activation ............ 4 1.2.1 Lipopolysaccharide as model for sepsis and systemic immune activation ........................ 7 1.2.2. Sepsis induced by Gram positive bacteria ........................................................................ 8 1.3. Chronic central nervous system infection with Toxoplasma gondii........................... 9 1.4. Immune cells of the central nervous system .......................................................... 10 1.5 Learning and memory............................................................................................. 14 1.6. Functional and structural synaptic plasticity of hippocampal neurons .................... 16 1.7 Crosstalk between the immune and central nervous system .................................. 18 1.8 Alzheimer´s disease ............................................................................................... 19 1.9 Alzheimer´s disease and innate immune response ................................................. 21 1.10. Aim of study ........................................................................................................ 24 2. MATERIAL AND METHODS ....................................................................................... 25 2.1. Material ................................................................................................................. 25 2.1.1. Chemicals ........................................................................................................................25 2.1.2. Immune stimulatory reagents ..........................................................................................25 2.1.3. Buffers and solutions .......................................................................................................25 2.1.4. Antibodies ........................................................................................................................26 2.1.5. Mouselines ......................................................................................................................27 2.1.5.1. APP/PS1 transgenic mice ............................................................................................27 2.1.5.2. NLRP3 knockout mice ..................................................................................................27 2.2. Methods ................................................................................................................ 27 2.2.1. Genotyping of APP/PS1 transgenic mice ........................................................................27 2.2.2. Immune stimulation with endotoxic bacterial cell wall components ................................29 2.2.3. Infections with Toxoplasma gondii ..................................................................................29 2.2.4. Perfusion of the mouse brain and preparation of fixed brain slices ................................30 2.2.5. DiOlistic labeling of single neurons .................................................................................30 2.2.6. Imaging and analysis of DiI stained neurons ..................................................................31 2.2.7. Immunohistochemical analysis of immune response in the central nervous system ......32 2.2.8. Preparation and incubation of hippocampal acute slices ................................................32 2.2.9. Electrophysiological recordings .......................................................................................33 2.2.9.1. Input-Output curve ........................................................................................................33 2.2.9.2. Paired pulse facilitation................................................................................................ 34 2.2.9.3. Long-term potentiation................................................................................................. 34 2.2.10. Morris water maze navigation task ............................................................................... 35 2.19 Statistical analysis ............................................................................................................ 38 3. RESULTS ................................................................................................................... 39 3.1. Long-term effects of an immune stimulation with different kinds of LPS ................ 39 3.1.1. Influence of LPS E.coli and LPS S.typhimurium on neuronal morphology in aged mice40 3.1.2. Effects of LPS E.coli and LPS S.typhimurium on synaptic plasticity in old mice ........... 43 3.1.3. Influence of LPS E.coli and LPS S.typhimurium on learning behavior of aged mice ..... 45 3.1.4. Quantification of glial immune response in the brain of old immune stimulated mice.... 49 3.1.5. Influence of E.coli and S.typhimurium LPS on neuronal morphology in adult mice ....... 51 3.1.6. Comparison of the glial phenotype in young and old mice ............................................. 53 3.1.7. Changes in neuronal morphology upon LPS stimulation in aged APP/PS1 mice.......... 55 3.1.8. Synaptic plasticity in aged APP/PS1 mice 3 months after stimulation with LPS............ 58 3.1.9. Neuronal architecture of young APP/PS1 mice 3 months after stimulation with LPS .... 59 3.1.10 Changes in neuronal architecture of aged NLRP3 knockout mice after peripheral immune stimulation ................................................................................................................... 62 3.1.11 Synaptic plasticity of aged NLRP3 KO mice 3 months after stimulation with LPS ....... 64 3.2. Peripheral immune stimulation with components of Gram positive bacteria .......... 66 3.2.1. Long-term effects of PGN/LTA from S.aureus on neuronal morphology in aged mice .. 67 3.2.2. Enduring changes of PGN/LTA of S. aureus on synaptic plasticity in old mice ............. 69 3.2.3. Peptidoglycan/lipoteichoic acid treatment affects learning behavior of aged mice ........ 70 3.2.4. Influence of an immune stimulation with PGN/LTA on the glial immune response ....... 73 3.2.5. Long-term influences of PGN/LTA on neuronal
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