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標題: | 轉基因斑馬魚肌肉專一性表現Progranulin基因導致肌肉細胞肥大之分子機制研究 Molecular Mechanisms Study of Myocyte Hypertrophy in Transgenic Zebrafish with Muscle-specific Overexpression Progranulin Gene |
作者: | Chia-Hsuan Liao 廖嘉瑄 |
指導教授: | 吳金洌 |
關鍵字: | 肌肉生成調控因子,肌肉肥大,轉基因魚, progranulin,MRFs,hypertrophy,transgenic line,zebrafish, |
出版年 : | 2007 |
學位: | 碩士 |
摘要: | Progranulin(Pgrn) 被歸類為上皮組織生長因子的家族,具有調節系統發育、傷害復原等功能並且已知與數種癌症的發生有關。先前研究顯示,腹腔注射生長激素(GH)會誘導肝臟的Pgrn伴隨著IGF-1一起表現。已經證實GH-IGF-1系統在個體生長上具有關鍵的調節功能。而肌肉為魚體高比例組織。因此,我們藉由肌肉專一性的表現Pgrn來探討此生長因子在斑馬魚肌肉生長所扮演的角色。首先,從斑馬魚肝臟中複製pgrn cDNA。此cDNA 包含962氨基酸其中包含11個granulin次單元。從不同時間點分析基因表現量之結果顯示,pgrn表現於魚卵中具有母體效應而胚胎從6 somite 階段開始表現。位置上,在發育的過程pgrn基因的表現分布廣泛。 組織分布上,pgrn基因主要分布在肝,腸,腎和脾臟。 使用tol2 transposon系統, 我們建立肌肉專一性大量表現pgrn轉基因魚。相較於原生株,轉基因魚之體重和體長均顯著的增加。轉基因公母魚的促進體重比率分別為48.14%和 48.00%。在轉基因公母魚增長的體長比率各別為25.14% 和75.93%。接著以切片,HE染色的方式來分析肌肉細胞在型態上的變化。取二個月之斑馬魚肌肉組織分析各個肌肉細胞之平均面積,結果顯示相較於野生株(255.70 Progranulin (Pgrn) was considered as a family of epithelial tissue growth factor because of the potential functions displayed in the regulation of development, wound healing and progression of several cancer types. In our previous study, we found hepatic pgrn and IGF-1 was co-induced with growth hormone (GH) administration. As well known that GH-IGF1 axis plays a critical role in regulating of somatic growth and the majority of fish body is muscle tissue. Therefore, we attempt to prove the function of pgrn in zebrafish by muscle-specific over-expression approach. First, the pgrn cDNA was cloned from zebrafish liver cDNA. The cDNA contains an open reading frame encoding a peptide 962 amino acid residues (aa) that contains eleven granulin motifs. The temporal expression shown that pgrn gene possesses a maternal expression and initiated expression from 6-somites stages. The tissue distribution of pgrn gene was major contributed in liver, intestine, kidney and spleen. Using tol2 transposon system, a zebrafish transgenic line with muscle-specific over-expression of pgrn was established. The body weight and body length were increased significantly in transgenic line compared with that in wild type. The promoted ratio of body weight in transgenic male and female were 48.14% and 48.00%, respectively. The promoted ratio of body length in transgenic male and female were 25.14% and 75.93%, respectively. The morphology of muscle cells in sections were clarified by HE stain. The average size of muscle cells in a select area were increased significantly in transgenic line (345.14 |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28262 |
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