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博碩士論文 etd-0818109-153528 詳細資訊
Title page for etd-0818109-153528
論文名稱
Title
斑鰭飛魚(Cyselurus poecilopterus)骨骼肌之蛋白質體研究
Study on the Proteomics of Flyingfish (Cyselurus poecilopterus) Skeletal Muscle
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
56
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2009-07-27
繳交日期
Date of Submission
2009-08-18
關鍵字
Keywords
斑鰭飛魚、骨骼肌、蛋白質體
skeletal muscles, proteomics, Cyselurus poecilopterus, flyingfish
統計
Statistics
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中文摘要
飛魚受驚嚇時會迅速擺動尾鰭衝出水面,並張開特化的胸鰭作長距離的滑翔。此特化的胸鰭是靠著兩群肌肉控制伸展與收縮,這兩群的肌肉外觀均呈現淡紅色,但未經組織學的證實,且兩者生化特徵比較也尚無描述。本研究目的在於比較斑鰭飛魚胸鰭肌肉與軀幹白肌和紅肌在組織學和蛋白質體學上的差異,並試圖瞭解胸鰭肌肉與體側紅肌之差異,以及其功能。利用斑鰭飛魚作為實驗物種,我們發現,胸鰭肌肉肌纖維束面積介於白肌和紅肌之間,且具有較大面積的結締組織與些許的脂肪組織,所以飛魚的胸鰭肌肉可能不歸類為白肌或紅肌,並且胸鰭肌肉會行脂質代謝提供能量來維持鰭的伸展。利用蛋白質體學比較三種肌肉,其中肌肉收縮蛋白actin,myosin regulatory light chain,myosin light polypeptide,酵素蛋白isocitrate dehydrogenase,malate synthase,queuosine biosynthesis protein和兩個逆境蛋白 heat shock protein (HSP70 and HSP60) 的蛋白質含量在胸鰭肌肉高於白肌或紅肌。這些結果顯示飛魚的胸鰭肌肉可能兼行有氧和糖解的代謝功能,肌纖維型態應屬於中間型肌纖維的粉紅肌。
Abstract
Flying fish has specialized pectoral fins. When they are activated, they will rush out of the water, expand their pectoral fins and flap their caudal fin to glide. The pectoral fins are controlled by two groups of muscles in which the external appearance is pink. No histological investigations have been made on their muscles to verify whether they are red muscles. The purposes of this study were to compare the pectoral fin muscle, trunk white muscle and trunk red muscle by histological and proteome methods so as to understand if the pectoral fin muscles is red muscles and to infer their function. Cyselurus poecilopteins was used for this study, Result show that the sizes for the cross section of the pectoral-fin-muscle-fibers were between the white and red muscles, and a large amount of connective tissue and fat tissues are present in the space among the muscle cells. It is interpreted the pectoral fin muscles of flying fish might not belong to white muscle and red muscle, and they probably utilize lipid metabolism to provide enough energy for the gliding activates. The proteomic pages for the three muscle types were compared and differences were found in the muscle proteins: actin, myosin regulatory light chain, myosin light polypeptide; enzymes: isocitrate dehydrogenase, malate synthase, queuosine biosynthesis protein;stress proteins: heat shock protein (HSP70 and HSP60). Expressions of these proteins were high in the pectoral-fin muscles than in the white and red muscles. These results suggest that the flying fish’ pectoral-fin muscles may involve in the oxidative and glycolysis pathways, and the muscle fibers type maybe belong to an intermediate type of muscle fiber.
目次 Table of Contents
總目錄
章次 頁次
謝辭……………………………………………………………………………………i
中文摘要………………………………………………………………………………ii
英文摘要……………………………………………………………………………...iii
總目錄…………………………………………………………………………………v
表目錄………………………………………………………………………………..vii
圖目錄……………………………………………………………………………….viii
一、前言………………………………………………………………………………1
1.1 斑鰭飛魚簡介………………………………………………………...………1
1.2 行為…………………………………………………………………………...1
1.3 形態學…………………………………………………………………...……2
1.4 白肌與紅肌………………………………………………………………...…2
1.5 蛋白質體學…………………………………………………………...………3
1.6 研究目標…………………………………………………………………...…4
二、材料與方法………………………………………………………………………6
2.1 實驗魚種………………………………………………………………...……6
2.2 樣品製備…………………………………………………………………...…6
2.3 組織切片………………………………………………………………...……6
2.4 組織切片分析……………………………………………………………...…7
2.5 蛋白質純化……………………………………………………………...……7
2.6 蛋白質定量……………………………………………………………...……7
2.7二維電泳法 (two-dimensional gel electrophoresis) ……………………….…8
2.7.1 第一維 等電膠集電泳 Isoelectric focusing electrophresis (IEF) …….8
2.7.2 平衡 Equilibration…………………………………...…………………9
2.7.3 第二維 聚丙醯胺膠體電泳 SDS-PAGE…………………….........…10
2.7.4 Coomassie blue G-250染色…………………………………...……….11
2.8 Gel image analysis………………………………………………………...…12
2.9 In gel digestion…………………………………………………………….…12
2.10 MALDI-TOF操作……………………………………………………….…12
2.11 MSCOT辨認未知蛋白……………………………………………………..13
三、結果…………………………………………………………………....…………14
3.1 胸鰭肌肉織型態………………………………………………………….…14
3.2 肌肉組織的差異………………………………………………………….…14
3.3 肌肉間蛋白質的表現………………………………………………….……15
四、討論………………………………………………………………………………17
4.1 肌肉組織差異性分析………………………………………………….……17
4.2 蛋白質差異性分析……………………………………………………….…17
五、結論……………………………………………………………………....………22
六、參考文獻…………………………………………………………………………23
表……………………………………………………………………………………..28
圖……………………………………………………………………………………..34
個人履歷……………………………………………………………………………..45
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