The role of astrocytic RFX4_v3 in demyelination [thesis]

Paap Franziska
Introduction 1.1 Multiple sclerosis 1.1.1 Symptoms, clinical course and diagnosis 1.1.2 Pathology of MS 1.1.3 Pathogenesis 1.1.4 The role of astrocytes in the pathogenesis of MS 1.1.5 The cuprizone mouse model 1.2 Regulatory factor X4: Member of the protein family regulatory factor X 1.2.1 RFX4_v3: The brain specific variant of RFX4 1.2.2 RFX4_v3 deficiency leads to non-communicating congenital hydrocephalus 1.2.3 Potential target genes and interacting partners of RFX4_v3 1.3 Hydrocephalus
more » ... Congenital hydrocephalus 1.3.2 Subcommissural organ and Reissner fiber glycoproteins 1.4 Aims 2 Materials & methods 2.1 Materials 2.1.1 Reagents 2.1.2 Solutions, buffers and cell culture media 2.1.3 Enzymes/ proteins 2.1.4 Kits 2.1.5 Monoclonal antibodies for flow cytometry 2.1.6 Primary antibodies for immunohistochemistry and fluorescence immunohistochemistry 2.1.7 Secondary antibodies for immunohistochemistry and fluorescence immunohistochemistry 2.1.8 Oligonucleotide primers 2.1.9 TaqMan Expression Assays 2.1.10 Consumables 2.1.11 Technical devices 2.1.12 Software 2.1.13 Tissue from MS patients 2.2 Animals 2.3 Mouse strains 2.3.1 Housing 2.4 Methods 2.4.1 Genotyping 2.4.2 PCR for the analysis of RFX4 transcript variants in MS 2.4.3 Cuprizone mouse model 2.4.4 Histology 2.4.5 Luxol Fast Blue -Periodic Acid Schiff (LFB-PAS) staining 2.4.6 Hematoxylin and eosin (H&E) staining 2.4.7 Immunohistochemistry and fluorescence immunohistochemistry 2.4.7.1 Immunohistochemistry 2.4.7.2 Fluorescence immunohistochemistry 2.4.8 Morphometry and data aquisition 2.4.9 Isolation of astrocytes from newborn mice 2.4.10 In vitro migration assay 2.4.11 Quantitative PCR analysis 2.4.11.1 Isolation of RNA I would like to offer my special thanks to Koki, Martina, Alexandra, Pia, Sarah and Thorben for their friendship and for the emotional and constant support. Further, I am grateful to Alexandra and Martina for proofreading of this thesis. Very personal thanks to my parents Maria and Wolfgang and my siblings Kathrin and Dominik for always supporting me. Finally, I want to thank my boyfriend Tjebbe for his emotional support, motivation, trust and endless patience.
doi:10.53846/goediss-5062 fatcat:ivcxlhvqsfexjol6nkyaxkdfxe