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研究生:王昭華
研究生(外文):Chao-Hua Wang
論文名稱:台灣土雞產蛋能力與脂肪細胞功能之探討
論文名稱(外文):The laying performance and adipose function in Taiwan country chicken
指導教授:陳珠亮
指導教授(外文):Chu-Liang Chen
學位類別:碩士
校院名稱:東海大學
系所名稱:畜產與生物科技學系
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:2006
畢業學年度:95
語文別:中文
論文頁數:97
中文關鍵詞:台灣土雞脂肪細胞功能產蛋能力
外文關鍵詞:Taiwan country chickenadipose functionlaying performance
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為了瞭解雞隻產蛋能力與脂肪細胞功能的關係,本試驗以25週齡國立中興大學由同源雞群長期選育具有高產蛋能力的L2品系台灣雞隻與低產蛋能力的B品系台灣雞隻做為試驗材料,取其腹脂分析比較兩品系間脂肪細胞功能與前脂肪細胞分化能力。由甘油-3-磷酸去氫酶(GPDH)活性分析的結果顯示,L2品系的脂質合成能力高於B品系;脂質分解分析方面,L2品系脂肪細胞的脂解能力亦明顯高於B品系。而以相對定量反轉錄聚合酶連鎖反應分析脂肪組織中的脂肪細胞與Stromal-Vascular 細胞(S-V 細胞)的基因表現,發現在S-V 細胞,許多與脂肪細胞功能有關的基因,都以L2品系高於B品系,如:PPARγ、C/EBPβ、LPL、ADRP和adipoQ,顯示L2品系S-V 細胞有較多已分化的脂肪細胞。在成熟脂肪細胞方面,L2品系與脂質代謝相關基因,如:FAS、perilipin和Glut1的表現都明顯高於B品系,說明L2品系成熟的脂肪細胞具有較高的脂質合成能力與脂質分解的能力,同時也具有較高的葡萄糖吸收能力。另一方面、S-V 細胞誘發分化處理後,經oil-Red O脂質染色與GPDH活性分析結果,顯示分化與脂質堆積能力,以L2品系高於B品系;而在分化過程中,L2品系的PPARγ、aP2及adipoQ基因的表現明顯高於B品系,顯示L2品系的前脂肪細胞分化能力高於B品系。進一步發現,S-V 細胞經分化為脂肪細胞後,培養於不含葡萄糖的培養液中,L2與B品系細胞的GPDH活性並無差異,說明L2細胞對葡萄糖的敏感度高於B品系細胞。以上試驗結果顯示,高產蛋率L2品系雞隻的前脂肪細胞分化能力較高,且具有較高的脂質合成與脂解作用,說明雞隻具活躍的脂肪細胞功能與前脂肪細胞分化能力,可能較能有效地提供產蛋時所需脂質,而有助於產蛋的表現。
In order to understand the relationship between chicken laying performance and adipocyte functions, this study used 25 week-old chickens of the L2 and B2 strain that displays high and low laying performance trait, respectively, as the model. Analysis of adipocyte function and preadipocyte differentiation activity was carried out using cells derived from the abdominal adipose tissue. Based on glycerol-3-phosphate dehydrogenase (GPDH) activity, higher lipid synthesis activity was observed in L2 than B. Higher lipolysis activity was also detected in L2 than B adipocyte. The results of relative quantitative reverse transcription-polymerase chain reaction (RT-PCR) indicated that higher mRNA levels of genes involved in adipocyte functions, such as PPARγ, C/EBPβ, LPL, ADRP and adipoQ, were recorded in the stromal-vascular (S-V) cells prepared from L2 than B. The results indicate that L2 has more mature adipocytes among its S-V cells than B. Moreover, fatty acid synthesis, lipolysis, and glucose intake are more active in these mature adipocytes of L2 than B, as higher mRNA levels of lipid metabolism-related genes, including FAS, perilipin and Glut1, are detected in mature adipocytes prepared from L2 than B. Furthermore, S-V cells were treated for adipocyte differentiation. The results of oil-Red O staining and GPDH activity assay showed that higher ratios of the S-V cells of L2 have the potential to differentiate into adipocyte than those of B, since higher mRNA levels of genes participating in adipocyte differentiation, such as PPARγ, aP2, glut1 and adipoQ, were detected during adipocyte differentiation of the S-V cells in L2 than B. Finally, lower GPDH activity was detected in mature adipocyte developed from the S-V cells of L than those of B, suggesting that L2 cells are more sensitive to glucose concentration than B cells. Overall, preadipocytes in the abdominal adipose tissue of L2 strain apparently have higher adipocyte differentiation potential as well as greater lipid synthesis and lipolysis activity than those of B strain. It is likely that higher activity potential in adipocyte function and differentiation may provide the energy for egg production more efficiently to facilitate reproduction performance.
目錄
摘要 4
前言 5
文獻檢討 7
壹、台灣養雞事業之發展 7
貳、禽類脂質代謝 8
參、禽類產蛋能力與脂質代謝的關係 9
肆、脂肪組織的種類與功能 11
伍、脂肪細胞研究模式 13
一、脂肪細胞 13
二、體內試驗(in vivo) 13
三、體外試驗(in vitro) 14
四、細胞培養之建立 15
五、脂肪細胞分化 17
陸、脂肪細胞分化代謝之基因表現 18
一、Peroxisome proliferator-activated receptors γ(PPARγ) 18
二、Lipoprotein lipase(LPL) 20
三、CCAAT/Enhancer-binding protein-β(C/EBPβ) 21
四、Adipocyte differentiation-related protein(ADRP) 23
五、Fatty acid synthase(FAS) 24
六、Adipocyte fatty acid binding protein(A-FABP) 25
七、Glucose transporter 1(Glut1) 26
八、Adiponectin、adiponectin recepter1和adiponectin recepter2 28
九、Vimentin 29
十、Perilipin 31
材料與方法 33
壹、材料 33
(一) 動物: 33
(二) 試驗材料: 33
貳、方法 34
(一) 動物飼養 34
(二) 細胞培養 34
(三) 甘油-3-磷酸去氫酶活性分析 (GPDH activity assay) 34
(四) 脂解作用分析 (Lipolysis assay) 36
(五) 總RNA的萃取 36
(六) 相對定量反轉錄聚合酶連鎖反應(relative quantitative reverse transcription polymerse chain reaction; RT-PCR) 36
(七) Oil-red O染色法與定量 38
(八) 統計分析 38
結果與討論 40
試驗一、雞前脂肪細胞分化培養方法之建立 40
一、不同血清來源對雞前脂肪細胞分化之影響 40
﹙一﹚ 胎牛血清與雞血清對雞前脂肪細胞分化形態之影響 40
﹙二﹚ 不同濃度之胎牛血清與雞血清對雞前脂肪細胞分化脂質堆積之影響 41
﹙三﹚ 不同濃度胎牛血清與雞血清對前脂肪細胞分化相關基因表現之影響 41
二、雞前脂肪細胞之分化培養 42
(一) 雞前脂肪細胞分化培養之GPDH活性分析 42
(二) 雞前脂肪細胞分化培養過程相關基因表現之分析 42
試驗二、不同產蛋能力雞隻脂肪細胞脂質代謝能力分析 43
一、脂肪組織GPDH活性之比較 43
二、脂肪細胞脂解作用之分析 43
試驗三、不同產蛋能力雞隻脂肪細胞基因表現 45
一、脂質代謝相關基因表現之比較 45
二、細胞油滴結構相關基因表現之比較 46
三、動物能量代謝相關基因表現之比較 47
四、前脂肪細胞分化相關基因表現之比較 47
試驗四、不同產蛋能力雞隻前脂肪細胞分化培養之脂肪功能表現比較 49
一、前脂肪細胞分化外觀之比較 49
二、前脂肪細胞分化脂質堆積之比較 49
三、脂肪細胞脂質堆積與培養液葡萄糖濃度關係之比較 50
(一) 葡萄糖濃度對脂肪細胞形態影響之比較 50
(二) 葡萄糖濃度對脂肪細胞GPDH活性之比較 51
試驗五、不同產蛋能力雞隻前脂肪細胞分化過程之相關基因表現之比較 52
一、分化相關基因表現之比較 52
二、脂質代謝相關基因表現之比較 53
三、細胞油滴結構相關基因表現之比較 54
四、動物能量代謝相關基因表現之比較 55
結論 57
參考文獻 79
英文摘要 97
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