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日本鰻血管生成素(Angiopoietin)-1基因選殖與血管新生相關因...

國立高雄大學生物科技研究所

日本鰻血管生成素(Angiopoietin)-1基因選殖與血管新生相關因子在紅體組織及卵巢表現調控之研究 = Cloning of Angiopoietin-1 gene and study of the regulation on angiogenic factors in the rete mirabile and ovary in Japanese eel (Anguilla japonica)

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	Cloning of Angiopoietin-1 gene and study of the regulation on angiogenic factors in the rete mirabile and ovary in Japanese eel (Anguilla japonica)
作者:	張宸瑋,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	2009[民98]
面頁冊數:	178面圖、表 : 30公分;
標題:	紅體
標題:	Catfish Pituitary Exacts (CPE)
附註:	指導教授：施能朗、黃永森
附註:	參考書目：面
摘要註:	血管新生為動物體內一受到高度調控之生理功能。在外在環境刺激，如: 傷口癒合、缺氧等情形下，刺激血管內皮細胞活化多種血管生長因子表現。其中血管生成素-1在血管新生後期與其受器結合後，可活化PI3K-Akt等多種訊息傳遞路徑，誘導血管成熟相關功能表現，故推測血管生成素-1與血管之生長分化和成熟有關。本研究以斑馬魚血管生成素-1 mRNA序列為標的來設計引子，選殖日本鰻(Anguilla japonica)血管生成素-1的部份片段(812 bps)。其胺基酸序列與斑馬魚血管生成素-1有80%的相似性，採用選殖出之片段設計引子並運用聚合連鎖反應分析鰻魚組織及活體外細胞之血管生成素-1表現量，以日本鰻之Angiopoietin-1(Ang-1) mRNA序列探討血管生成素-1與相關血管生長因子在此類原始脊椎動物出生後，發育成熟至可降海繁殖階段的生理功能。在活體(in vivo)實驗有三部分，一、取樣體重100克至400克之日本鰻，並分析其紅體組織的相關血管生長因子表現量，發現體重300克至400克組別，血管生成素-1 mRNA表現量最高，顯示在體型較大之日本鰻其血管生成素-1表現較多。推測在體型較大鰻魚之血管新生較趨近成熟。二、日本鰻每週注射鯰魚腦下垂體萃取液催熟(Catfish Pituitary Exacts, CPE)，於第0週、第3週、第6週、第9週後取紅體組織分析。血管生成素-1在第六週表現達高峰且與紅體生長呈正相關。推測在催熟後血管生成素-1表現逐漸達高峰。三、日本鰻每週注射鯰魚腦下垂體萃取液催熟，於第0週、第3週、第6週、第9週後取卵巢分析，並由其中隨機挑出十隻合併注射睪固酮，發現僅注射腦下垂體萃取液組別其相關血管生長因子與GSI值呈負相關，但合併注射睪固酮9針之組別與注射腦下垂體萃取液9針之組別，GSI值有明顯提升且各血管生長因子表現亦增加，顯示睪固酮可刺激卵巢內血管生長因子表現而促進卵巢發育。在活體外(in vitro)實驗部分，日本鰻初代紅體細胞以不同濃度PPAR agonists、內皮細胞生長補充劑(ECGs)、類固醇、生長因子、金屬離子等處理特定時間後，調查紅體細胞內血管生成素-1及相關生長因子受調控情形。發現PPAR γ agonist刺激VEGF表現; ECGs刺激Ang-1、VEGF與FLK表現;鋅離子亦可刺激Ang-1、VEGF與FLK等因子表現; 鈷離子只能刺激VEGF表現;睪固酮刺激Ang-1但抑制VEGF;黃體酮刺激Ang-1與FLK;雌二醇刺激Ang-1、FLK與podocalyxin;胰島素刺激Ang-1與VEGF表現; IGF-1刺激Ang-1、Eph與podocalyxin; bFGF刺激Ang-1與VEGF表現。了解日本鰻的紅體與卵巢血管新生過程中，血管生成素-1等因子所扮演角色，期望未來能將血管生成素-1製成重組蛋白以改善日本鰻人工繁殖中，由於血管生長不完全而導致卵巢發育不良與幼鰻發育缺陷的問題。 Angiogenesis is a highly regulated physiological process in animals. Among the different angiogenic factors, angiopoietin-1 combines with its receptor to induce many signal transduction pathways related to vessel maturation including PI3K-Akt pathway in late phase of angiogenesis, which maintain endothelial cells survival through anti-apoptosis function, recruit pericytes supplements and migration and make compact interaction with vessel tubes. Angiopoietin-1 is highly associated with vessel maturation. We designed degenerate primers from Angiopoietin-1 of zebrafish and cloned a part of nucleic sequence from Anguilla japonica (812 bp), and its amino acid sequence has 80% similarity with Angiopoietin-1 of zebrafish. I used this segment to design specific primers to analyze Angiopoietin-1 expression in tissue or cells of Anguilla japonica with polymerase chain reaction (PCR), investigating physiological function of Angiopoietin-1 and relating angiogenic factors during development after birth in such a primitive vertebrate. In in vivo experiments - I sampled Anguilla japonica that weight range from 100g to 400g and then analyzed Angiopoietin-1 and relating angiogenic factors expression in the rete mirabile. And I found that Angiopoietin-1 express uppermost in the group that weight range from 300g ~ 400g, it suggests Angiopoietin-1 express higher in the larger eels and their angiogenesis is near to maturation. 2. Rete mirabile from Anguilla japonica injected with catfish pituitary exacts (CPE) once every week, and sampled on the week one、week three、week six、week nine. After analyzing, we found Angiopoietin-1 increase gradually and express uppermost on the sixth week and decrease on the ninth week. We suggest Angiopoietin-1 express higher in the late phase of injecting CPE to promote vessel maturation in rete mirabile. 3. Japanese eel injected with CPE once every week, and sampled on the week one、week three、week six、week nine. The expression of Ang-1 and other angiogenic factors decrease with increasing GSI index, but the GSI index increase obviously and angiogenic factors expression rescue in the group that extra injecting testosterone. It reveals that testosterone may stimulate angiogenic factor activity and promote ovary growth. In in vitro, rete mirabile cells were treated with various PPAR agonists、endothelial cell growth supplement (ECGs)、steroid、growth factor、metal ions, and we investigate how Angiopoietin-1 and relating angiogenic factors being regulated. We expect to find out the characters of Angiopoietin-1 and other angiogenic factors during angiogenesis in the rete mirabile and ovary from Anguilla japonica. Furthermore, we apply to solve the problem in eel artificial reproduction due to incomplete blood vessel development.

日本鰻血管生成素(Angiopoietin)-1基因選殖與血管新生相關因子在紅體組織及卵巢表現調控之研究 = Cloning of Angiopoietin-1 gene and study of the regulation on angiogenic factors in the rete mirabile and ovary in Japanese eel (Anguilla japonica)
張, 宸瑋

日本鰻血管生成素(Angiopoietin)-1基因選殖與血管新生相關因子在紅體組織及卵巢表現調控之研究 = Cloning of Angiopoietin-1 gene and study of the regulation on angiogenic factors in the rete mirabile and ovary in Japanese eel (Anguilla japonica) / 張宸瑋撰 - [高雄市] : 撰者, 2009[民98]. - 178面 ; 圖、表 ; 30公分.
指導教授：施能朗、黃永森參考書目：面.
紅體Catfish Pituitary Exacts (CPE)

日本鰻血管生成素(Angiopoietin)-1基因選殖與血管新生相關因子在紅體組織及卵巢表現調控之研究 = Cloning of Angiopoietin-1 gene and study of the regulation on angiogenic factors in the rete mirabile and ovary in Japanese eel (Anguilla japonica)
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