Comparative proteomic analysis of the mouse ovary in response to GnIH and GnRH: An in vitro approach

Anushree Dave, Michael D. Powell, R. Sridaran, K. Tsutsui, A. Krishna
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Abstract

The present study conducted proteomic analysis to identify ovarian proteins induced by GnRH/GnIH in comparison with untreated control mice. We used methods of 2-dimentional gel electrophoresis and LC-MS/MS for protein identification. Out of the 25 differentially expressed spots from each of the GnRH/GnIH treated ovaries we were randomly selected 18 protein spots identified by LC-MS/MS. The proteins 14-3-3, prohibitin and superoxide dismutase (SOD1) identified by LCMS/MS were further verified by Western blotting, which confirmed that GnRH/GnIH-induced changes in expression of 14-3-3, prohibitin and SOD1 were similar to the observations of proteomics. These selected proteins, 14-3-3, prohibitin and SOD1, were further have been the correlation with various ovarian activities, such as cell proliferation, survival or markers of apoptotic. Out of 18 identified GnRH-induced proteins, 9 showed up-regulations whereas other 9 showed down-regulations. This finding suggests that GnRH acts both by stimulating and inhibiting expression of protein. Interestingly, in the GnIH-induced ovary, 15 out of 18 proteins were significantly down-regulated suggesting that, GnIH acts mainly by inhibiting the expression of proteins involved in ongoing ovarian activities. Thus, the present study indicates many of the identified GnRH/GnIH-induced proteins are potentially linked to ovarian follicular development, atresia, steroidogenesis and cancer.
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小鼠卵巢对GnIH和GnRH反应的比较蛋白质组学分析:体外方法
本研究通过蛋白质组学分析鉴定了GnRH/GnIH诱导的卵巢蛋白,并与未治疗的对照组进行了比较。采用二维凝胶电泳和LC-MS/MS方法进行蛋白鉴定。从每个GnRH/GnIH处理卵巢的25个差异表达点中,我们随机选择18个通过LC-MS/MS鉴定的蛋白点。LCMS/MS鉴定的14-3-3、prohibitin和超氧化物歧化酶(SOD1)蛋白进一步通过Western blotting进行验证,证实GnRH/ gnih诱导14-3-3、prohibitin和SOD1的表达变化与蛋白质组学观察结果相似。这些选择的蛋白,14-3-3,prohibitin和SOD1,进一步与卵巢的各种活动,如细胞增殖,存活或凋亡标志物相关。在鉴定的18个gnrh诱导蛋白中,9个表达上调,而另外9个表达下调。这一发现表明GnRH通过刺激和抑制蛋白质的表达来起作用。有趣的是,在GnIH诱导的卵巢中,18种蛋白中有15种显著下调,这表明GnIH主要通过抑制参与卵巢活动的蛋白的表达来起作用。因此,本研究表明,许多已鉴定的GnRH/ gnih诱导蛋白可能与卵巢卵泡发育、闭锁、类固醇生成和癌症有关。
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