碩士 / 國立臺灣海洋大學 / 生物科技研究所 / 95 / PCTAIRE protein kinases belong to a subfamily of cdc2-related serine/ threonine protein kinases. The PCTAIRE protein kinase family, which have masses of 50-60 kDa, consist of three isoforms in mammals and are ubiquitously expressed in the brain. PCTAIRE-1 has been recently reported to play a possible role in regulating neurite outgrowth (Graeser et al., 2002). In addition, PCTAIRE-1 has been discovered to interact with 14-3-3 proteins (Sladeczek et al., 1997). Expression of different isoforms of 14-3-3 GFP-fusion protein via a neuron-specific HuC promoter in zebrafish embryos indicated that only 14-3-3-beta3 could promote neurite outgrowth. Based on this result, we suggest that PCTAIRE-1 may be a regulator of neurite outgrowth. We first cloned zebrafish PCTAIRE-1 gene, and demonstrated that this gene was expressed in the zebrafish brain by whole mount in situ hybridization. In order to understand the function of this gene in CNS development, we designed four mutant forms of zebrafish PCTAIRE-1. Besides, we overexpressed both wild type and mutant forms of zPCTAIRE-1-GFP fusion proteins driven by neuronal specific HuC promoter by injecting into zebrafish embryos at one-cell stage. The amount of the green fluorescent cells in zebrafish, either injected with wild type or mutant zPCTAIRE-1-GFPs, were less than that of HuC-GFP control observed at 48 hpf. However, co-injection of pHuC-Bcl-2-GFP with wild type or mutant zPCTAIRE-1 into zebrafish embryos could maintain the survival of some neuron cells. It is suggested that the anti-apoptotic Bcl-2 gene could prevent the neuronal apoptosis caused by zPCTAIRE-1. Based on these results, we suggest that zPCTAIRE-1 possibly regulates the apoptosis of zebrafish nervous system. On the other hand, co-injection of pHuC-14-3-3-Beta3-GFP with zPCTAIRE-1 wild type or mutant forms into zebrafish embryos showed that only zPCTAIRE-1-K167R could suppress the neurite outgrowth promoted by the 14-3-3-beta3 isoform. Therefore, we suggest that the kinase activity of zebrafish PCTAIRE-1 may regulate the z14-3-3-beta3 isoform and subsequent neurite outgrowth.
| Identifer | oai:union.ndltd.org:TW/095NTOU5111011 |
| Date | January 2007 |
| Creators | Shin-Hua Tsai, 蔡欣樺 |
| Contributors | Chang-Jen Huang, 黃銓珍 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 72 |
Page generated in 0.002 seconds