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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/34640

Title: hif2α、birc5a與birc5b基因弱化抑制斑馬魚胚胎內neurod基因群表現
Inhibition of neurod genes by hif2α、birc5a and birc5b morpholino knockdown in zebrafish embryos
Authors: Wei-Yi Chen
陳瑋毅
Contributors: NTOU:Institute of Bioscience and Biotechnology
國立臺灣海洋大學:生物科技研究所
Keywords: 斑馬魚;視網膜;中樞神經
neurod;hif;birc5a;birc5b
Date: 2012
Issue Date: 2013-10-07T02:44:43Z
Abstract: 在脊椎動物胚胎發育過程中,basic-helix-loop-helix (bHLH)轉錄因子有著重要作用,而低氧濃度誘導因子(Hypoxia-inducible factors,HIFs) 會在缺氧環境下負責調控生理狀態。NeuroD為一種屬於bHLH的轉錄因子,表現在CNS、neuronal cell等部位,含有調控細胞週期、控制神經元分化的功能。在斑馬魚發育過程中, hif2α弱化後,會降低細胞凋亡抑制蛋白survivin (birc5a 與birc5b)的表現,造成細胞凋亡以及神經分化受到影響,此外也會抑制neurod表現。但是hif2α弱化後對於其他 neurod基因的影響及這些基因是否參與視網膜以及神經的發育至今尚不清楚。本實驗主要為觀察在hif2α、birc5a 與birc5b弱化的情況下對於各種neuroD members基因的影響。實驗中利用全覆式原位雜交染色觀察班馬魚胚胎內各種neurod members的表現位置及狀況,再利用morpholino抑制hif2α、birc5a 或birc5b表現後觀察對各neurod member基因的影響,實驗結果顯示在胚胎發育過程中,neurod與neurod4 的表現最明顯,主要表現在視網膜與中樞神經細胞中,其次是neurod2在視網膜與中樞神經的表現,而neurod6a及-6b的表現較微弱,並僅出現在中樞神經的部位。hif2α弱化後,發現除了neurod6b之外,各neurod members表現受到抑制,表示在斑馬魚胚胎發育當中hif2α對於其他neurod members也具有調控的作用。birc5a或是-5b的弱化亦會降低neurod6b之外各neurod members的表現,但是降低之幅度較hif2α弱化輕微。實驗結果顯示在胚胎發育過程中,hif2α、birc5a/5b與neurod members間有著緊密的關聯,而這些基因在視網膜細胞的分化過程中所扮演的功能將留待後續探討。
Basic-helix-loop-helix (bHLH) transcription factor plays an important role in the process of vertebrate embryonic development, and the hypoxia -inducible factors (HIFs) play a key role to adapt physiological functions during hypoxic stress. NeuroD is a basic helix loop helix transcription factor that is widely expressed in CNS and neuronal cells. NeuroD regulates cell cycle and control neuronal differentiation. Knock down hif2α diminished survivin (birc5a and birc5b) expression and caused severe neuronal apoptosis and impaired neuronal differentiation. In addition, blocking hif2α translation also repressed neurod transcription. However, it is unclear whether other neurod members are also influenced by hif2α suppression. Here I found that the neurod and neurod4 are the major neurod members expressed in retina and CNS during development. A less amount of neurod2 transcript is also observed in retina and CNS regions during development. neurod6a and neurod6b are weakly expressed in specific CNS regions during development. Knockdown hif2α expression suppressed all members of neurod expression, except neurod6b, Knockdown birc5a and birc5b expression also decreased all members of neurod mildly. In summary, this study revealed that neurod, neurod4 and neurod2 are expressed in retina and CNS during development and their expressions were influenced by hif2α, birc5a and birc5b depletion.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0019936037
http://ntour.ntou.edu.tw/handle/987654321/34640
Appears in Collections:[生命科學暨生物科技學系] 博碩士論文

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