Roy Shyamal K, Wang Cheng, Mukherjee Anindit, Chakraborty Prabuddha
Departments of OB/GYN and Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, NE, USA.
Methods Mol Biol. 2012;825:151-71. doi: 10.1007/978-1-61779-436-0_12.
Follicular development commences with the formation of primordial follicles, which begins with the differentiation of pluripotent ovarian somatic cells into early granulosa cells and their apposition to the oocytes in the egg nest. The process of primordial follicle morphogenesis and factors affecting the formation and development of primordial follicles can be examined in vitro using fetal ovaries in organ culture. The functions of candidate genes involved in primordial folliculogenesis can be examined using siRNA or shRNA, which can knockdown specific mRNA targets at specific time points. Here, we describe the organ culture protocol for fetal hamster ovary with GPR30 siRNA as an example. The method to morphologically analyze follicular development is also discussed.
卵泡发育始于原始卵泡的形成，原始卵泡的形成始于多能性卵巢体细胞分化为早期颗粒细胞，并与卵母细胞在卵母细胞巢中并置。原始卵泡形态发生过程以及影响原始卵泡形成和发育的因素可以在体外使用器官培养的胎儿卵巢进行研究。参与原始卵泡发生的候选基因的功能可以使用siRNA或shRNA进行检测，它们可以在特定时间点敲低特定的mRNA靶点。在这里，我们以用GPR30 siRNA处理的胎儿仓鼠卵巢为例描述器官培养方案。还讨论了卵泡发育形态学分析方法。
