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牛排卵前卵泡的基因表达谱分析:促性腺激素峰及前列腺素依赖性上调可能与排卵过程相关的基因

Gene expression profiling of bovine preovulatory follicles: gonadotropin surge and prostanoid-dependent up-regulation of genes potentially linked to the ovulatory process.

作者信息

Li Qinglei, Jimenez-Krassel Fermin, Ireland James J, Smith George W

机构信息

Laboratory of Mammalian Reproductive Biology and Genomics, Michigan State University, East Lansing, Michigan 48824-1225, USA.

出版信息

Reproduction. 2009 Feb;137(2):297-307. doi: 10.1530/REP-08-0308. Epub 2008 Nov 7.

Abstract

The molecular mechanisms of ovulation and luteinization have not been well established, partially due to lack of a comprehensive understanding of functionally significant genes up-regulated in response to an ovulatory stimulus and the signaling pathways involved. In the present study, transcripts increased in bovine preovulatory follicles following a GnRH-induced LH surge were identified using microarray technology. Increased expression of 368 and 878 genes was detected at 12 (368 genes) and 20 h (878 genes) following GnRH injection. The temporal, cell specific and prostanoid-dependent regulation of selected genes (ADAM10, DBI, CD36, MTSS1, TFG, and RABGAP1) identified from microarray studies and related genes (ADAM17 and AREG) of potential significance were also investigated. Expression of mRNA for DBI and CD36 was simultaneously up-regulated in theca and granulosa cells (GC) following the LH surge, whereas temporal regulation of ADAM10, MTSS1, TFG, and RABGAP1 was distinct in the two cell compartments and increased granulosa TFG and RABGAP1 mRNA were prostanoid dependent. AREG mRNA was increased in theca and GCs at 12 and 24 h following GnRH injection. ADAM17 mRNA was increased in theca, but reduced in GCs 24 h following GnRH injection. The increased ADAM17 and AREG mRNA were prostanoid dependent. ADAM10 and ADAM17 protein were increased specifically in the apex but not the base of preovulatory follicles and the increase in ADAM17 was prostanoid dependent. Results reveal novel information on the regulation of preovulatory gene expression and suggest a potential functional role for ADAM10 and ADAM17 proteins in the region of follicle rupture.

摘要

排卵和黄体化的分子机制尚未完全明确,部分原因是对排卵刺激后上调的功能重要基因以及相关信号通路缺乏全面了解。在本研究中,利用微阵列技术鉴定了GnRH诱导LH峰后牛排卵前卵泡中增加的转录本。在GnRH注射后12小时(368个基因)和20小时(878个基因)检测到368个和878个基因的表达增加。还研究了从微阵列研究中鉴定出的选定基因(ADAM10、DBI、CD36、MTSS1、TFG和RABGAP1)以及具有潜在重要性的相关基因(ADAM17和AREG)的时间、细胞特异性和前列腺素依赖性调节。LH峰后,DBI和CD36的mRNA表达在卵泡膜细胞和颗粒细胞(GC)中同时上调,而ADAM10、MTSS1、TFG和RABGAP1在两个细胞区室中的时间调节不同,颗粒细胞中增加的TFG和RABGAP1 mRNA是前列腺素依赖性的。GnRH注射后12小时和24小时,AREG mRNA在卵泡膜细胞和颗粒细胞中增加。GnRH注射后24小时,ADAM17 mRNA在卵泡膜细胞中增加,但在颗粒细胞中减少。增加的ADAM17和AREG mRNA是前列腺素依赖性的。ADAM10和ADAM17蛋白在排卵前卵泡的顶端而非基部特异性增加,ADAM17的增加是前列腺素依赖性的。结果揭示了排卵前基因表达调控的新信息,并表明ADAM10和ADAM17蛋白在卵泡破裂区域具有潜在的功能作用。

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