The mouse follicle microenvironment regulates antrum formation and steroid production: alterations in gene expression profiles.
The mouse follicle microenvironment regulates antrum formation and steroid production: alterations in gene expression profiles.
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DOI：
10.1095/biolreprod.108.071142
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发表时间：
2009-03
影响因子：
3.6
通讯作者：
Shea LD
中科院分区：
文献类型：
--
作者：
West-Farrell ER;Xu M;Gomberg MA;Chow YH;Woodruff TK;Shea LD
卵泡发生是一个协调的过程，调节发育的基因很难在体内研究。体外培养系统允许在发育过程中评估单个卵泡，从而使基因表达模式在卵泡发育过程中得到监测。小鼠多层次卵泡（直径150-180 μm）在不同物理性质的三维基质中培养8天。在体外卵泡生长期间，监测卵泡形成和类固醇产生，并分离mRNA。检测与卵泡发育状态相关的基因（Star、Cyp11a1、Cyp17a1、Hsd3b1、Cyp19a1、Fshr、Lhcgr、Aqp7、Aqp8、Aqp9、Hif1a）的表达水平。形成腔并产生适当水平的雌激素和孕酮的卵泡，Star、Aqp7、Aqp8和Hif1a的表达不变，在培养第8天Cyp19a1的表达增加了34倍，在培养第6天和第8天Lhcgr升高。然而，健康的卵泡，但没有形成腔或产生适当水平的类固醇，显示Star， Aqp7， Aqp8和Hif1a水平增加，Cyp19a1在培养第8天增加15倍，Lhcgr水平直到培养第8天才升高。本研究首次利用海藻酸盐水凝胶的个体培养对基因表达进行了时间分析，该基因表达与卵泡发育过程中的生长和甾体生成有关，并确定了健康卵泡和发育不良卵泡的表达模式。
Folliculogenesis is a coordinated process, and the genes that regulate development are difficult to investigate in vivo. In vitro culture systems permit the assessment of individual follicles during development, thereby enabling gene expression patterns to be monitored during follicle development. Mouse multilayered secondary follicles (150–180 μm in diameter) were cultured in three-dimensional matrices of varying physical properties for up to 8 days. During this period of follicle growth in vitro, antrum formation and steroid production were monitored, and mRNA was isolated. The expression levels of genes (Star, Cyp11a1, Cyp17a1, Hsd3b1, Cyp19a1, Fshr, Lhcgr, Aqp7, Aqp8, Aqp9, and Hif1a) were measured and correlated to follicle developmental status. Follicles that developed an antrum and produced appropriate levels of estrogen and progesterone had unchanging expression of Star, Aqp7, Aqp8, and Hif1a, a 34-fold increase in Cyp19a1 expression at day 8 of culture, and elevated Lhcgr at days 6 and 8 of culture. Follicles that were healthy but did not form an antrum or produce appropriate levels of steroids, however, demonstrated increasing levels of Star, Aqp7, Aqp8, and Hif1a, a 15-fold increase in Cyp19a1 at day 8 of culture, and Lhcgr levels were not elevated until day 8 of culture. This study provides the first temporal analysis of gene expression using individual culture in alginate hydrogels, which correlates growth and steroidogenesis during follicle development and identifies expression patterns in healthy and in developmentally disadvantaged follicles.