Effects of the glutamic acid decarboxylase (GAD) inhibitor semicarbazide and anti-GAD autoantibodies-containing immunoglobulin G on neuronal network activity within the motor cortex [Elektronische Ressource] / vorgelegt von Christina Ingrid Holfelder

Effects of the glutamic acid decarboxylase (GAD)inhibitor semicarbazide and anti-GADautoantibodies-containing immunoglobulin G onneuronal network activity within the motor cortexDissertation zur Erlangung des Doktorgrades derNaturwissenschaftenFakultät für Biologie der Ludwig-Maximilians-UniversitätMünchenvorgelegt vonChristina Ingrid HolfelderApril 20101. Gutachter: Prof. Mark Hübener2.: Prof. Hans StrakaTag der Einreichung: 29.04.2010Tag der mündlichen Prüfung: 04.08.2010ContentsAbbreviations VII1 Introduction 11.1 γ-Aminobutyric Acid (GABA) . . ....................... 11.2 Glutamic Acid Decarboxylase (GAD) . .................... 21.3 Semicarbazide (SMC) . . ............................ 41.4 Anti-GAD Autoantibodies (Anti-GAD AAbs) . ................ 61.5 Introduction to the Technique of Intrinsic Optical Signal (IOS) Recording . 101.6 Goals of this PhD Thesis . . . . . ........................122 Materials and Methods 152.1 Solutions and Drugs ...............................152.2 Purification of SPS-IgG and Control IgG....................172.2.1 Purification of IgG with Affinity Chromatography . .........172.2.2 Buffer Exchange . . . . .182.2.3 Quantification of IgG Concentration . . ................182.3 Detection of Anti-GAD AAbs in Purified SPS-IgG . . . . . .........192.3.1 Purification of Mouse Brain Proteins . .192.3.2 Western Blots192.3.3 GAD-Dot . . ...............................202.4 Preparation of Mouse Brain Slices . . . ............