2015-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/642753摘要：本研究是以多產、發育快速、容易進行胚胎觀察的斑馬魚作為實驗動物模式，其神經系統發育在卵受精後只需五天即可初步完成結構發育。我們初步發現 inaa (α-internexin同源基因)的核糖核酸會廣泛的表達在斑馬魚胚胎和成體的腦部及視網膜中，然而沒有相關研究探討此神經中間絲蛋白在斑馬魚中樞神經系統發育的功能。本研究目的是將探討斑馬魚 inaa 蛋白在發育中斑馬魚中樞神經系統之表現與分布狀況，接著將更進一步研究 inaa 在中樞神經系統發育時的功能。在第一年計畫中，我們將製作多株抗體標定 inaa蛋白，並確認其效力與專一性，用免疫組織染色確認 inaa 在發育中樞神經系統之表現與分布狀況。在第二年，我們將利用設計好的 Mopholino 抑制此蛋白的表現，以原位雜交、免疫組織染色和電子顯微技術去探討 inaa 在斑馬魚中樞神經系統發育時的功能。本計畫研究結果可應用於以斑馬魚模式的中樞神經系統研究，且可以釐清 inaa在斑馬魚中樞神經系統發育時的功能。經由了解 inaa 在斑馬魚中樞神經系統所扮演的角色，將有助於進一步釐清神經中間絲蛋白可能參與相關神經疾病的機轉。 <br> Abstract: In this study, we use the zebrafish as our animal model, which is characterized by productiveness, rapid development and easy in-situ observation of embryonic development. As early as 5 day after fertilization, all major components of the zebrafish central nervous system (CNS) are developed. Our preliminary data showed that the mRNA of inaa, an ortholog of the mammalian α-internexin, was widely expressed in the brain as well as retina of zebrafish from the embryonic to adult stages. The functional role inaa played in the development of zebrafish CNS has not been extensively studied. The aims of this proposal are to illustrate the timing and spatial distribution of inaa protein in the developing zebrafish CNS and then to analyze the function of inaa during the development of zebrafish CNS. In the first year, we will generate a specific polyclonal anti-inaa antibody and confirm the protein distribution in the developing zebrafish CNS by immunohistochemistry. In the second year, we will knockdown the expression of inaa by using morpholinos and then analyze the function of the inaa during the development of zebrafish CNS by in situ hybridization, immunohistochemistry and electron microscopy. This project result can be applied to the study of zebrafish CNS and will clarify the role of inaa in the development of zebrafish CNS. Furthermore, to understand the function of inaa in the CNS will offer us deep insights into the mechanism of neuronal intermediate filaments in the neurological diseases.α-internexin斑馬魚中樞神經系統神經發育α-internexinzebrafishcentral nervous systemneural development