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研究生:陳玟鴒
研究生(外文):Wen-Ling Chen
論文名稱:雌性素受體在乙型轉型生長因子促進卵泡促素誘發隙裂接合蛋白表現與功能的角色
論文名稱(外文):Role of Estrogen Receptor in Follicle-Stimulating Hormone and Transforming Growth Factor beta 1 Upregulation of Connexin Gap Junction
指導教授:黃娟娟黃娟娟引用關係
指導教授(外文):Jiuan-Jiuan Hwang
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生理學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:英文
論文頁數:46
中文關鍵詞:雌性素受體乙型轉型生長因子卵泡促素隙裂接合蛋白
外文關鍵詞:estrogen receptorTransforming Growth Factor beta 1Follicle-Stimulating HormoneConnexin Gap Junction
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Connexin (Cx) 所構成的隙裂接合(gap junction)在發育及器官功能上扮演著相當重要的角色。而Cx43為卵巢顆粒細胞中最為主要的隙裂接合組成,若卵巢中的Cx43基因被剃除,則小鼠的濾泡發育將只成長至早期,且卵子也會有發育遲緩的現象。有報導指出雌激素可增加子宮肌膜層的Cx43表現,但作用機制仍不明確。除此之外,我們先前的實驗指出,若是抑制Cx43通道蛋白或是雌激素受體 (ER),將減低FSH和TGFbeta1所增進大鼠卵巢顆粒細胞的類固醇生成。因此,本論文研究目的為探討在大鼠卵巢顆粒細胞中,ER是否媒介FSH和TGFbeta1所促進Cx43蛋白生成及隙裂接合功能,以及那一類型的ER對於此一效應是較為主要的。另外亦研究TGFbeta1作用是否透過TGF�涀eta第一型受體以加強FSH增進Cx43生成及隙裂接合功能。西方墨點法分析結果顯示FSH和TGFbeta1增加磷酸化Cx43 (Cx43-P1及Cx43-P2)的含量,而此效應為前處理ER拮抗劑 (ICI-182780)或ERalpha拮抗劑 (MPP)所抑制,但卻不受ERbeta拮抗劑 (PHTPP) 前處理的影響。而此一抑制情形也可在FSH和TGFbeta1所增進隙裂接合功能中觀察到。此外,我們發現前處理TGFbeta第一型受體抑制劑 (SB431542)可抑制FSH及TGFbeta1所誘發的Cx43生成及隙裂接合功能。綜合上述,本研究結果証實在大鼠卵巢顆粒細胞中,ERalpha以及TGFbeta第一型受體媒介FSH和TGFbeta1所增進Cx43生成及隙裂接合功能。
Connexin gap junctions play important roles in development and organ functions. Cx43 is the predominant connexin present in granulosa cells of ovarian follicles. In Cx43 knockout mice, folliculogenesis did not proceed beyond the primary follicle stage, and oocyte growth was retarded. Estrogen was reported to upregulate Cx43 expression in rat uterine myometrium; however, the mechanism remains unclear. In addition, our previous studies have shown that inhibition of Cx43 intercellular junction or estrogen receptor (ER) suppressed the FSH and TGFbeta1-promoted steroidogenesis in rat ovarian granulosa cells. Therefore, this study was to investigate whether ER mediates FSH and TGFbeta1 upregulation of Cx43 gap junction in rat ovarian granulosa cells, and which ER isoform is responsible for the induction. In addition, we determined whether TGFbeta1 acts through its type I receptor (TGFbetaR-I) to enhance FSH stimulation of Cx43 gap junction. Using western blot analysis, we demonstrate that FSH and TGFbeta1 greatly increased the level of phosphorylated Cx43, and this was reduced by pretreatment with ER antagonist (ICI-182780) and ERalpha selective antagonist (MPP), but not ERbeta selective antagonist (PHTPP). Moreover, the suppressive effect of ER antagonists was also seen in FSH and TGFbeta1-increased gap junction communication. Additionally, we show that TGFbetaR-I blocker (SB431542) suppressed the FSH and TGFbeta1-increased Cx43 protein level and gap junction communication. In summary, this study suggests that ERalpha and TGFbetaR-I mediate FSH and TGFbeta1-induced intercellular communication in rat ovarian granulosa cells.
English abstract 1
Chinese abstract 2
Introduction 3
A.Ovarian folliculogenesis 3
B.Role of follicle-stimulating hormone (FSH) in the ovary 5
C.Role of transforming growth factorbeta (TGFbeta) in the ovary 7
D.Role of estrogen in the ovary 9
E.Role of gap junction in the ovary 11
F. Objective 14
Materials and Methods 15
A. Materials 15
B. Animals 15
C. Cell culture and treatment 16
D. Immunoblot analysis of Cx43 17
E. Gap junction communication: scrape-loading dye transfer assay 18
F. Statistics 18
Results 19
A. Effects of ER antagonists on FSH and TGFbeta1-induced Cx43 level and gap junction communication 19
B. Involvement of TGFbetaR-I in TGFbeta1 enhancement of FSH- induced Cx43 level and gap junction function 20
Discussion 30
References 33
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