第一型类胰岛素生长因子基因转殖动物之产制及分析.docVIP

第一型類胰島素生長因子基因轉殖動物之產製及分析Generation and Analysis of Insulin-like growth factor-1 Transgenic Animals 研究生：謝政宏Cheng, Hung Hsieh 指導教授：蘇弘毅Hung, Yi Su 【摘要】 第一型類胰島素生長因子 insulingrowth factor 1; IGF-1 在活體的作用已有研究明確指出，類胰島素生長因子包括IGF-1和IGF-2，均具有媒介荷爾蒙GH 之功能，循環中的IGF-1主要是由肝臟所合成及分泌，且局部組織亦產生IGF-1，二者分別具媒介GH的內及自體/旁分泌功能 Daughaday, 1989 。目前之研究顯示，局部產生的IGF-1在乳腺的發育及乳汁的生產較循環之IGF-1重要，且IGF-1除可媒介GH之作用治療人類之生長不全；亦可生長；而由於其與胰島素之結構相似，亦可治療糖尿病。因此本論文的研究目的，擬將外源性的IGF-1，藉一乳腺專一性之啟動子將其導引至動物之乳腺細胞，而期望基因轉殖增加之外源IGF-1增加細胞之增生，導致泌乳量激增。本實驗室先前已有報告顯示，以牛之 -LA（ -lactalbumin 乳腺專一性啟動子下游接一羊之IGF-1 cDNA，產製出之IGF-1基因轉殖鼠，其泌乳量與同窩非轉殖鼠比較，在泌乳期可顯著的（p＜0.05）增加，但該報告僅分析比較3隻基因轉殖與3隻非基因轉殖鼠，而本論文擬產製較多基因轉殖鼠，以確認先前之結果；且以同樣之α-LA-IGF-1構築基因產製基因轉殖乳山羊，並使用各種年齡的母山羊將當做胚的者與供者，ontrolled internal drug release CIDR ，以superovulation 與發情同期化 estrus synchronization 之處理，肌肉注射，黃體荷爾蒙 progesterone 、孕馬血清激性腺素 pregnant mares serum gonadotropin 、人類絨毛膜激性腺素 human chorionic gonadotropin, CG 、卵胞刺激荷爾蒙 follicle stimulating hormone FSH 及激黃體素uteinizing hormone, LH ，CIDR移除後約24小時，的交配，手術由輸卵管收集胚。在完成核顯微注射後，將2~3個胚轉移至者輸卵管側，以產生基因轉殖親代動物，，。關鍵詞:IGF-1、基因轉殖乳山羊、原核顯微注射The somatomedin hypothesis proposes that the insulin-like growth factors, IGF-1 and IGF-2, mediate action of the growth hormone GH, somatotropin . Circulating IGF-1 is mainly synthesized and secreted by the liver but it is also produced locally in tissues. The hepatic-derived and local tissues produced IGF-1, mediates the endocrine and autocrine/paracrine action of GH, respectively Daughaday, 1989 . Existing data suggest that locally produced IGF-1 is more important to the development of mammary gland and secretion of milk compared to the systemic IGF-1. As IGF-1 mediates action of GH, it can be used for the treatment of insufficient growth, but also normalise the growth excess, which is excluded by GH treatment. As the chemical structure of IGF-1 is similar to the insulin, it can be used in the diabetic patients. In this study, we set out to produce IGF-1 transgenic mice and goats in which overexpression of ectopic IGF-1 may