斑馬魚Atox1對於血管形成和造血功能是扮演重要的角色

中山醫學大學機構典藏 CSMUIR > 醫學科技學院 > 醫學檢驗暨生物技術學系暨碩士班 > 博碩士論文 > Item 310902500/10147

Please use this identifier to cite or link to this item: https://ir.csmu.edu.tw:8080/ir/handle/310902500/10147

Title:	斑馬魚Atox1對於血管形成和造血功能是扮演重要的角色 The Zebrafish Atox1 is Crucial for Vasculogenesis and Hematopoiesis.
Authors:	吳嘉斌 Wu, Chin-Bin
Contributors:	中山醫學大學:醫學檢驗暨生物技術學系;蔡淦仁
Keywords:	Atox1;銅離子;血管新生;造血作用;原位雜交技術 Copper;Atox1;vasculogenesis;hematopoiesis;in situ hybridization
Date:	2014
Issue Date:	2014-12-10T04:11:36Z (UTC)
Abstract:	銅離子為生物體的必須微量元素之一，參與各種生理性的過程，包括細胞呼吸作用、抗氧化防禦機制、神經物質合成等。而Atox1是含量最多的銅離子結合蛋白，並負責運送銅離子到高爾基氏體參與的運輸網絡內。先前的研究證實，當老鼠體內缺乏銅離子，會導致其胚胎產生缺陷、以及卵黃囊中的血管異常，但Atox1在此過程中所扮演的作用仍未清楚。為了解銅離子及Atox1於血管新生、甚至造血作用之間的關連性，在本研究中我們利用注射反義股(antisense)的Atox1核?酸到轉基因斑馬魚(fli-1:EGFP)胚胎，進行基因抑制實驗來觀察斑馬魚在胚胎發育上的影響。我們的結果發現，這些Atox1被抑制的斑馬魚它們的 ISV有錯誤配對、DLAV及PAV的異常，我們發現與Atox1的缺損有高度的關連性，與銅離子缺乏無關。O-dianisidine染色，觀察紅血球，也發現斑馬魚的造血也受到影響。利用Real-time PCR及原位雜交技術，研究其Atox1所缺陷胚胎的發育機制。我們發現一些血管和造血的基因，包括flk、scl、lmo2等；以及血管新生的基因，包括vegfa、flt、angiopoietin-2等都會受到影響。綜合以上的證據，所以我們提出Atox1在早期胚胎發育中於血管新生與造血作用扮演了關鍵性的角色。 Copper is an essential trace element for organisms, involved in various physiological processes including respiration, antioxidant defense and neurotransmitter biosynthesis, and many others. The Atox1 is the most abundant copper-binding protein and responsible for copper delivery to the secretory proteins within the trans-Golgi network. Previous study has demonstrated the copper deficiency results in embryonic defects and yolk sac vasculature abnormalities in mouse; however, the role of Atox1 in these processes is yet to be defined. To study the role of copper and Atox1 in vasculogenesis and hematopoiesis during embryo developments, zebrafish (fli-1:EGFP) was used to prepare Atox1 knockdown fish by antisense morpholino injection. Using O-dianisidine staining to examine the hematopoiesis of RBC, zebrafishes with disorganized pattern of the intersegmental vessels (ISV), the malformation of dorsal longitudinal anastomotic vessel (DLAV) and parachordal vessels (PAV) were observed, and strongly associated with the loss of Atox1, but not copper. Real-time PCR and in situ hybridization technique were taken to investigate the development of atox1- deficiency embryos. Our data revealed that angiogenic and hematopoietic gene, including flk, scl, and lmo2, and the vasculogenesis genes, including vegfa, flt, and angiopoietin-2 were affected in Atox1-deficiency embryos. Together all these evidences, we have proposed a crucial role for copper-chaperon Atox1 in vasculogenesis and hematopoiesis during embryonic development.
URI:	https://ir.csmu.edu.tw:8080/ir/handle/310902500/10147
Appears in Collections:	[醫學檢驗暨生物技術學系暨碩士班] 博碩士論文

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