Functional identification of zebrafish Nogo-relatedgenes in vitro and in vivo / 斑馬魚抑制脊椎再生蛋白Nogo之功能研究

碩士 / 國立臺灣大學 / 生化科學研究所 / 92 / In mammal, Nogo is a myelin-associated inhibitor of axon regrowth, which is involved in the regeneration of injured axons in central nervous system (CNS). Despite papers regarding the function and regulation of mammalian Nogo proteins increased rapidly, no report about Nogo gene in zebrafish has been proposed. Here we reported three homologous genes to RTN family in zebrafish, termed z-Nogo-B, -C1, and –C2. As in mammal, these transcripts come from the alternative splicing and promoter usage. So far, fish is the only vertebrate lacking of Nogo-A expression.

Our results indicated that cellular localization of zNogo-B was restricted in ER, while zNogo-C1 and zNogo-C2 in Golgi complex. When expressed zNogo-B in neuroblastoma cell line, the cellular localization of zNogo-B will switch to the ctyoplasm including the extending neurite. Knockdown of zNogo-B with morpholino antisense oligonucleotides caused neurite outgrowth, neuronal cell survive and somatic abnormalities. These results provide the functional evidence of z-Nogo-B and zNogo-C2 in zebrafish neuronal development.

The inhibitory pathway of Nogo-A is mediated by the binding with Nogo receptor. There were four likely Nogo receptors in zebrafish. To knockdown two of the four likely Nogo receptors in zebrafish, we could obtain the similar morphant as zNogo-B-Mo. They might be the receptor of the zNogo-B.

Our data revealed that the distribution of zNogo proteins in cell line and zebrafish embryos’ neuronal or glial system. By knocking down the zNogo-B expression could promote neurite outgrowth and neuronal cell survive in zebrafish embryos. Furthermore, while we knock down 2 of 4 Nogo receptors, we could obtain the similar phenotypes to Nogo-B morphant. Suggesting both 2 receptors might influence the neuron development in zebrafish embryos or act as receptors to zNogo-B.

Identiferoai:union.ndltd.org:TW/092NTU05103012
Date January 2004
CreatorsChien-Ming Lin, 林建銘
ContributorsChang-Jen Huang, 黃銓珍
Source SetsNational Digital Library of Theses and Dissertations in Taiwan
Languagezh-TW
Detected LanguageEnglish
Type學位論文 ; thesis
Format43

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