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系統識別號 U0026-0812200911161884
論文名稱(中文) 在腎臟發育的過程中Focal adhesion complex 蛋白調控之探討
論文名稱(英文) Regulation of Focal Adhesion Complex Proteins in Developing Kidney
校院名稱 成功大學
系所名稱(中) 生理學研究所
系所名稱(英) Department of Physiology
學年度 92
學期 2
出版年 93
研究生(中文) 蔡靜儀
研究生(英文) Ching-Yi Tsai
學號 s3690106
學位類別 碩士
語文別 英文
論文頁數 64頁
口試委員 口試委員-呂佩融
口試委員-陳鴻震
召集委員-沈孟儒
指導教授-湯銘哲
中文關鍵字 焦點黏著蛋白  腎臟發育 
英文關鍵字 kidney development  FAK 
學科別分類
中文摘要 在我們過去的實驗中, 已知道軟性的基質會去抑制細胞表現 FAK、paxillin 及talin 等等不同的焦點黏著複合蛋白。然而,這樣的現象對於生理上的重要意義卻不曾被探討過。在個體發育及成熟的過程中, 完整的FAK 仍然持續存在於腦部,心臟,肺及肝臟。相較之下, 我們發現到在胚胎發育至第十九天開始, 腎臟組織的FAK 會被分解100、90 及85 kDa 的N 端片段,隨後完整的FAK 則會在出生後完全消失。而其他的焦點黏著蛋白的表現量,例如paxillin、p130cas 及細胞內蛋白c-Src 也會隨著腎臟發育的過程中下降。為了確定是否基質軟硬度會影響腎臟中FAK 的表現,我們利用從三個月大的老鼠腎臟中所萃取出來的近端小管細胞來作初級培養。當初級近端小管細胞培養在一般的及覆蓋一層膠元蛋白的培養皿時,細胞會表現出焦點黏著複合蛋白。然而,將細胞培養在膠元蛋白凝膠則會抑制細胞再表現焦點黏著複合蛋白。在腎臟發育過程中或者是將初級進端小管細胞培養在不同軟硬的基質上時,FAK 的mRNA 表現並沒有改變,此現象顯示應是經由轉錄後修飾的機制來調控FAK 的表現量。在腎臟的切片染色實驗中,我們觀察到在胚胎時期時,FAK 的染色主要位在間質細胞,但是腎元卻沒有表現。而出生後,在腎臟皮質幾乎沒有FAK 的染色而主要表現在腎臟髓質內側上皮細胞。有趣的是在腎臟成熟的過程中,細胞核的FAK 染色非常顯著,暗示了85 kDa 的FAK 片段改變位置至細胞
核的角色。另外,在腎臟成熟的過程中,腎臟的髓質內N 端FAK 片段中的Y397, Y407 及Y577 位置有高量的磷酸化表現。綜合以上所述,本篇研究證明了在腎臟發育的過程中,基質的軟硬可能參與了調控焦點黏著複合蛋白的表現。而在腎臟成熟的過程中,sumoylation 的FAK可能是導致FAK 片段被磷酸化且將其轉換位置至細胞核的原因。

英文摘要 We previously showed that substratum rigidity down-regulated the expression of focal adhesion complex proteins such as FAK, paxillin and talin. However, the physiological significance of this finding has not been explored. Intact FAK remained constant levels in brain, heart, lung and liver throughout development and maturation. In contrast, we found that FAK was initially degraded into 100 kDa, 90 kDa and 85 kDa N-terminal fragments from embryonic 19.5 day and subsequently the intact
FAK was completely absent after birth in kidney. Other focal adhesion complex proteins such as paxillin, p130cas and cytosolic protein c-src were
also down-regulated during kidney development. To determine whether substratum rigidity affect expression of FAK, We employed primary culture of renal proximal tubular cells freshly isolated from 3-month-old rat kidney. Proximal tubular cells cultured on normal dish and collagen gel-coated dish exhibited focal adhesion complex proteins. However, collagen gel prevented the re-expression of focal adhesion complex proteins in primary culture. The expression level of FAK mRNA was not
altered in developmental kidney or in primary proximal tubular cells cultured on different rigidity, suggesting that the regulation of expression of FAK was mediated by post-transcriptional mechanism. Immunohistochemical study showed that FAK immunostaining were present in interstitial cells, but lost in developing nephrons in embryonic kidney. After birth, FAK immunostaining were completely absent in renal
cortex and expressed mostly in epithelial cells in inner medulla. Interestingly, nuclear localization of FAK immunostaining was quite prominent in mature kidney medulla, indicating the role of 85 kDa species in nuclear translocation. In addition, the phosphorylation at Y397, Y407 and Y577 of N-terminal FAK remained high in renal medulla during kidney maturation. Taken together, this study demonstrated that the rigidity might be involved in down-regulation of focal adhesion complex protein in developing kidney. Sumoylation of FAK may be the cases of phosphorylation of N-terminal FAK and translocation of N-terminal FAK
into nucleus of medullar tubular cells during kidney maturation.

論文目次 Abstract… … … … … … … … … … … … … … … Ⅰ
中文摘要… … … … … … … … … … … … … … … Ⅲ
誌謝… … … … … … … … … … … … … … … … … Ⅳ
Content… … … … … … … … … … … … … … … … Ⅴ
Figure content… … … … … … … … … … … … … Ⅵ
Introduction… … … … … … … … … … … … … … 1
Materials and methods… … … … … … … … … … … 8
Results… … … … … … … … … … … … … … … … 17
Discussion… … … … … … … … … … … … … … … 26
References… … … … … … … … … … … … … … … 32
Figures
作者簡歷
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