臺灣博碩士論文加值系統

斑馬魚的種種優點使此種模式生物已經成為實驗室中普遍的研究物種，也適合用來進行 (large-scale mutagenesis screen)。在斑馬魚中大量的突變分析是利用ENU(N-ethyl-N-nitrosourea)這個突變劑來使斑馬魚產生大量的突變，ENU是個烷化劑 (alkylating agent)，會作用在公魚的精母幹細胞(spermatogonial stem cell)，轉移它的乙烷基至DNA的氧或氮自由基上造成點突變。此研究對四種經過ENU注射而造成mutant的斑馬魚X-liver、Y-liver、Snake1及Snake2進行了型態的分析，發現四種mutant在5 dpf時皆因魚鰾萎縮而沉在底部不會游動，其中四種mutant的眼睛及頭部變的比較小；X-liver與Snake2的心臟有水腫的現象；而且Snake2的體軸有彎曲的情況；四種mutant的肝臟與腸道似乎都有發育不完全的情況。再利用Whole-Mount In Situ Hybridization的技術，用可以專一表現在各個消化性臟器的marker來評估mutant對斑馬魚消化性臟器發育上的影響，發現這四種mutant對肝臟、腸道、胰臟及胰島δ-細胞皆有影響。也利用多基因轉殖螢光魚LIP-αβ與mutant斑馬魚進行交配，利用螢光來觀察mutant對斑馬魚消化性臟器發育上的影響，發現在X-liver與Snake1的消化性臟器發出的螢光皆明顯的比wild type弱。這些結果顯示四種mutant皆對消化性臟器的發育造成嚴重程度不等的影響。

目錄.....I
表目錄.....III
圖目錄.....IV
中文摘要.....V
英文摘要.....VI
壹、序論.....1
一、動物模式物種 ─ 斑馬魚.....1
二、斑馬魚腸道、肝臟及胰臟的發育.....3
三、N-ethyl-N-nitrosourea (ENU)簡介.....7
四、指標基因(Marker Gene)簡介.....9
五、研究動機.....13
貳、實驗材料與方法.....15
一、實驗材料.....15
二、實驗方法.....22
參、實驗結果......33
一、Mutant斑馬魚的型態分析.....33
二、Whole-Mount In Situ Hybridization.....35
三、多基因轉殖螢光魚與Mutant斑馬魚.....42
肆、討論.....44
伍、參考文獻.....48
附表.....53
附圖.....55

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