Gene expression and Functional Identification of Zebrafish Myelin Basic Proteins
| Autor: | Yuan-Kai Tu, 杜元凱 |
|---|---|
| Rok vydání: | 2007 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 95 Myelin basic protein（MBP） is the major constitutional component in vertebrate myelin, which wraps axon rigidly and forms multilayered compact myelin sheath. The insulation nature of myelin sheath makes neuronal conduction impulse more efficient and faster. Many proteins which make up myelin including MAG, OMGP and NOGO, have been shown to be able to inhibit neurite outgrowth. In this study, we cloned zebrafish MBP genes and investigate whether fish MBP has similar function or not. First, by using bioinformatics and PCR technology, we cloned two zebrafish MBP isoforms: MBP1 and MBP2, and then analyzed the temporal and spatial expression pattern of MBP1 and MBP2 during zebrafish development by whole-mount in situ hybridization. MBP1 and MBP2 had similar expression pattern, but MBP1 was expressed in earlier stage than MBP2. Furthermore, in order to investigate the roles of MBP1 and MBP2 in HBNF-induced neurite outgrowth in zebrafish embryos, we co-injected MBP1/MBP2 and HBNF into zebrafish embryos. It was found that the degree of neurite outgrowth was decreased, and this decrease effect could be rescued by co-injecting with a neuroprotective factor, EPO. However, the reduction level of neurite outgrowth caused by MBP1 and MBP2 are different. In addition, by virtue of MO knockdown technology, we found some zebrafish embryos manifested developmental delay and died the next day after injecting MBP1-MO, and some MBP1 MO-injected embryos showed abnormal shake behaviors. In the case of MBP2-MO knockdown, we found obvious defect in the notochord formation during embryonic development. Finally, we generated a transgenic zebrafish line expressing MBP-GFP, and we utilized these MBP transgenic fishes to confirm above results of microinjection. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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