Molecular Mechanisms of Ovarian Follicular Activation

卵巢卵泡激活的分子机制

• 批准号: 6828485
• 负责人: JOANNE E. FORTUNE
• 金额: $ 10万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6828485
• 关键词: animal tissue; corpus luteum; developmental genetics; female; fertility; gene induction /repression; genetic mapping; graafian follicles; granulosa cell; in situ hybridization; ovary; reproductive development; subtraction hybridization

DESCRIPTION (provided by applicant): In female mammals most oocytes reside in primordial follicles in a resting stage. Little is known about the mechanisms that regulate the movement of these follicles into the growing pool (follicle activation). The pool of resting primordial follicles is a resource, yet untapped, that could be exploited as a source of material to provide alternative methods for alleviating infertility in women and propagating valuable domestic animals and endangered species. It is likely that follicular activation and growth in vivo are regulated by a delicate balance among various factors, both stimulators and inhibitors. To date, efforts to determine the signals that initiate follicle growth have consisted of testing individual "candidates" to determine if they promote or inhibit follicle activation. Although progress has been made using this approach, that progress has been slow. We seek support to develop a complementary approach, based on recent advances in molecular techniques, to determine specific genes that are turned on or off during the activation of follicles. In Specific Aim 1, suppressive subtractive hybridization (SSH) will be used to test the hypothesis that specific genes are turned on (or off) during follicle activation. Candidate genes will be determined by comparing freshly isolated pieces of bovine ovarian cortex (highly enriched for primordial follicles) with cortical pieces cultured for two days (in which 90% of follicles have activated and become primary). Cattle provide an excellent model for human follicular development and SSH are ideally suited for the detection of rare and novel sequences in two closely related cell types/tissues. Specific Aim 2 is designed to test candidate genes identified in Specific Aim 1 for a potential role in follicle activation by using in situ hybridization to localize differentially expressed sequences in freshly isolated vs. cultured bovine cortical pieces. At the end of two years, we expect to have identified a number of candidate genes that will provide important clues to the genetic regulation of follicle activation. The results generated by the proposed experiments will "open the door" to many applications, since it will be the first (to our knowledge) molecular and genetic approach to the regulation of follicle activation. Elucidation of these fundamental mechanisms has practical implications for the development of new contraceptive technologies and alleviation of infertility.

描述（由申请人提供）：在女性哺乳动物中，大多数卵母细胞都位于原始卵泡中。关于调节这些卵泡向生长池运动的机制知之甚少（卵泡激活）。静息原始卵泡的池是一种资源，但尚未开发，可以被用作材料来源，以提供减轻妇女不孕症并传播有价值的家畜和濒危物种的替代方法。卵泡激活和体内的生长可能受刺激剂和抑制剂之间的微妙平衡来调节。迄今为止，确定启动卵泡生长的信号的努力包括测试个体“候选”，以确定它们是促进或抑制卵泡激活的。尽管已经使用这种方法取得了进步，但进步却很慢。我们寻求支持基于分子技术的最新进展来开发一种互补方法，以确定卵泡激活期间打开或关闭的特定基因。在特定的目标1中，抑制性减法杂交（SSH）将用于检验以下假设：卵泡激活过程中特定基因被打开（或OFF）。候选基因将通过比较新鲜分离的牛卵巢皮质（高度富集原始卵泡）与培养两天的皮质碎片（其中90％的卵泡已激活并成为主要的皮质）来确定。牛为人类卵泡发育提供了出色的模型，SSH非常适合在两种密切相关的细胞类型/组织中检测稀有和新型序列。特定的目标2旨在通过使用原位杂交将差异表达的序列定位在新鲜分离的牛与培养的牛皮质碎片中，以测试特定目标1中鉴定出在卵泡激活中的潜在作用的候选基因。在两年结束时，我们预计将确定许多候选基因，这些基因将为卵泡激活的遗传调节提供重要的线索。所提出的实验产生的结果将“打开许多应用”，因为据我们所知，这将是第一种分子和遗传方法来调节卵泡激活。阐明这些基本机制对新的避孕技术和减轻不育的发展具有实际意义。