In vitro and in vivo analysis of the functional significance of Tenascin-R and -C, CD24 and Semaphorin3A for neural stem cell behaviour and axonal pathfinding in Mus musculus (L.) 1758 and Rattus norvegicus

................................................................................................................................................. 7 Zusammenfassung................................................................................................................................. 9 Abbreviations....................................................................................................................................... 11 General Introduction .......................................................................................................................... 15 Study one: Tenascin functions in neural stem cell behaviour ......................................................... 17 1.0. Introduction .................................................................................................................................. 17 1.1. Stem cells and their definitions .................................................................................................. 17 1.2. Stem cells in the development of the nervous system ................................................................. 18 1.2.1. Embryonic stem cells .......................................................................................................... 19 1.2.2. Adult stem cells ................................................................................................................... 20 1.3. Importance in gene therapy ....................................................................................................... 20 1.4. The TN family of ECM glycoproteins ........................................................................................ 23 1.4.1. Common structure of TNs................................................................................................... 24 1.4.2. The extracellular matrix glycoprotein TNC........................................................................ 27 1.4.3. Expression pattern of TNC in the nervous system .............................................................. 27 1.4.4. Functional characterization of TNC................................................................................... 28 1.4.5. TNC interactions................................................................................................................. 33 1.4.6. The extracellular matrixmolecule TN-R ............................................................................. 33 1.4.7. Expression pattern of TNR.................................................................................................. 34 1.4.8. Functional characterization of TNR................................................................................... 34 1.4.9. TNR interactions................................................................................................................. 37 1.5. Aims of study one ....................................................................................................................... 38 2.0 Materials and Methods ................................................................................................................. 39 2.1. Reagents, disposables, instruments............................................................................................ 39 2.2. Bacterial media .......................................................................................................................... 40 2.3. Buffers and stock solutions ........................................................................................................ 40 2.4. Molecular biological methods ................................................................................................... 41 2.4.1. Maintenance of bacterial strains ........................................................................................ 41 2.4.2. Production of competent bacteria ...................................................................................... 41 2.4.3. (Re-) Transformation of DNA into bacteria ....................................................................... 42 2.4.4. Purification of nucleic acids............................................................................................... 42 2.4.5. DNA agarose gel electrophoresis ....................................................................................... 44 2.4.6. Sequencing of DNA............................................................................................................. 44