臺灣博碩士論文加值系統

目前已知G蛋白信號調節因子（RGS）能夠直接與活化態的Gα結合，並刺激Gα次單元上的GTP酶的活性，加速GTP的被水解，可讓Gα蛋白變成不活化態。RGS蛋白依其結構與功能的不同可以區分為RGS1~RGS15家族，它們都具有一個120個胺基酸的保留結構區域，其功能主要是和活化態的Gα結合，負調節G蛋白的信號傳遞。許多RGS蛋白還具有其他的非RGS的結構區域，主要是能夠與其它的信號蛋白結合，來幫助調節G蛋白的信號傳遞過程。本論文主要探討斑馬魚的RGS7基因在不同的斑馬魚胚胎時期中是否有表現，及會表現在哪些組織部位；並且藉由注射專一性的反義寡核酸將斑馬魚胚胎RGS7基因剔除，觀察胚胎發育的影響。因此，利用RT-PCR以及全胚體原位雜合技術，結果發現在受精後0~96小時的斑馬魚胚胎中皆可偵測到RGS7的表現，其表現的位置是在斑馬魚胚胎的腦部、腹側主動脈、中期細胞團及體節等組織部位。再以顯微注射的技術將斑馬魚RGS7剔除，發現會干擾斑馬魚胚胎正常的心臟發育過程，影響心臟功能。結果顯示RGS7剔除後會使斑馬魚的心臟跳動微弱化、心包腔膨大以及心臟管狀化等現象。本研究證實RGS7蛋白於斑馬魚之早期心臟發育中扮演極重要之角色。

To date, it is well accepted that Regulator of G-protein signalling（RGS） proteins terminate signalling by accelerating the intrinsic G alpha-GTPase activity and recycling the G-protein complex back to its inactive GDP-bound heterotrimeric configuration. The RGS protein can be divided into 15 subfamilies（RGS1~RGS15） in accordance with the structure and the biological function. The RGS domain, a conserved 120-aa region exists in all RGS proteins, functions as a negative regulator through interacting with activated Gα subunit and turn off the signaling. In addition, RGS proteins contain other domains that can associate with other protein to enhance the G-protein signaling. In this study, we attempted to investigat the expression pattern and function of RGS7 in zebrafish during development. Using RT-PCR analysis, RGS7 was expressed from fertilization to 96 hours post-fertilization of Zebrafish. Expression of RGS7 was found in brain、aorta ventralis、intermediate cell mass and somite by whole-mount in situ hybridization. Microinjection of RGS7-specific antisense morpholino oligonucleotide into fertilized zebrafish embryos leaded to interfere normal heart development and the heart function. Phenotype such as weakness of heart beat, swollen pericardial cavity and abnormal heart tube formation was found in morphants. Taken together, there results suggest that RGS7 protein plays an important role in the early development of the zebrafish heart.

壹、 中英文摘要...................................1
貳、 縮寫檢索表...................................4
參、 序論.........................................5
肆、 研究動機....................................16
伍、 實驗方法與材料..............................17
陸、 實驗結果....................................39
柒、 討論........................................48
捌、 參考文獻....................................54
玖、 圖表與圖表說明..............................59

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