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研究生:范茗雯
研究生(外文):Ming-Wen Fan
論文名稱:黑鯛雌激素受器在生殖腺分化和性轉變的表現
論文名稱(外文):The expression of estrogen receptors in gonadal differentiation and sex change in the black porgy, Acanthopagrus schlegeli
指導教授:張清風張清風引用關係
指導教授(外文):Ching-Fong Chang
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:水產養殖學系
學門:農業科學學門
學類:漁業學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:73
中文關鍵詞:黑鯛先雄後雌雌激素雌激素受器性別分化性轉變
外文關鍵詞:black porgyprotandryestrogenestrogen receptorsex differentiationsex change
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雌激素藉由雌激素受器調控許多脊椎動物的生理作用,雌激素受器可分為細胞核上受器(erα、erβ1)及細胞膜上受器(gpr30),在魚類,另有一型魚類特有的erβ異構物,erβ2(erγ)。本研究目的主要探討雌激素受器於黑鯛性別分化及性轉變過程中基因表現模式。黑鯛生殖腺分化約為孵化後4個月至5個月,而erα、erβ1、erβ2於孵化後4個月表現量上升,於孵化後6個月下降,ers應和性別分化有關。雌激素受器表現量隨一年齡黑鯛精卵巢發育消長變動,erα、erβ2精巢組織內表現量高於卵巢組織;erβ1、gpr30卵巢組織內表現量高於精巢組織,顯示雌激素受器具有性別二相性。一年齡黑鯛去除精巢組織後提前至第二個繁殖季進行性轉變。erβ1於精巢切除後八個月(隔年一月)表現量升高,顯示erβ1參與卵巢組織的發育;gpr30的高表現量自切除後4個月(十月)持續至切除後8個月(隔年一月),顯示gpr30在卵巢發育與卵細胞成熟的過程中扮演重要的角色。二年齡黑鯛去除精巢後,輔以雌二醇(estradiol-17β, E2)投餵的結果指出erα、erβ2、gpr30進入繁殖季後,受到E2)的正向調節表現量提高。雌激素經由非基因型作用抑制hCG刺激生成的雄性素,gpr30於萎縮精巢內表現量高於活化精巢,推測gpr30在精巢萎縮的過程中扮演重要的角色。最後,雌激素受器在生殖腺分化、精卵巢組織發育、性轉變等過程中扮演極為重要的角色。
Estrogen mediates many physiological functions in vertebrata by estrogen receptors. Estrogen receptors include classical nuclear estrogen receptors (erα,erβ1) and membrane estrogen receptors (gpr30). There is second erβ isoform, erβ2 (erγ) in fish. The study aimed to investigate the role of estrogen receptors in the sexual differentiation and sex change. The gonadal sex differentiation in black porgy started at 120 to 150dph. Except gpr30, all of the genes expression incressed at 120dph and then decressed at 180 dph, indicating that estrogen receptors might be related to gonadal sex differentiation in black porgy. The expression of estrogen receptors was changed with different gonadal development stages. The gene expression of erα and erβ2 was higher in testis tissue than in the ovary tissue. On the contrary, the expression levels of erβ1 and gpr30 were high in the ovary tissue than in the testis tissue, suggesting that sex dimorphism existed in the gene expression of estrogen receptors. After the testis excision, 1+-year-old black porgy changed sex during the 2nd spawning season. The gene of erβ1 was expressed highly after 8 month testis excision (January), indicating that erβ1 was associated with the ovary tissue development. The gene of gpr30 was highly expressed after 4 month (October) to 8month (January) of testis excision, suggesting that gpr30 played an important role in the ovary development and oocyte maturation. After testis excision with E2-feeding in 2+-year-old fish, the results showed that E2 upregulated all the genes except erβ2 during spawning season. Estrogen decreased hCG-stimulated androgen synthesis in the testis via a nongenomic estrogen mechanism. The expression of gpr30 was higher in the regressed testis tissue than in the active testis, indicating that gpr30 was related to the regressed process in testis. In conclusion, estrogen receptors play important roles in gonadal sex differentiation, development, and sex change in black porgy.
壹、緒論 viii
1.雌激素在生殖腺發育的角色 1
2.環化酵素、雌激素與性別分化 2
3.雌激素受器 4
4.GPR30 6
5.研究源起 8
6.研究目的 9
貳、gpr30基因選殖與即時定量系統之建立 11
1.前言 11
2.實驗魚種 11
3.實驗設計 11
4.實驗方法 12
5.實驗藥品 20
6.結果 24
7.討論 25
參、黑鯛雌激素受器於性別分化與性別發育生殖腺組織之表現..........28

1.前言 28
2.實驗方法與設計 28
3.結果 30
4.討論 32
肆、黑鯛雌激素受器於人工誘發雌性發育時卵巢之變化 38
1.前言 38
2.實驗方法與設計 39
3.結果 43
4.討論 45
伍、 結論 55
參考文獻 57


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