Regulation of meiosis in mice

 描述（由申请人提供）：本申请的长期目标是阐明哺乳动物减数分裂的新分子网络，并开发用于体外减数分裂的哺乳动物细胞培养系统。减数分裂是生殖细胞特有的过程，包括配对联会、重组、联会消失和同源染色体分离。由减数分裂引起的遗传异常是人类出生缺陷、妊娠丢失和不育的主要原因。经历广泛重组的减数分裂染色质的结构和功能特性无疑是减数分裂的中心主题。尽管在了解减数分裂方面取得了快速进展，但染色质组织和减数分裂过程之间的复杂相互作用仍然在很大程度上未被探索。特别是，了解哺乳动物减数分裂的进展远远落后于其他模式生物的减数分裂研究，由于几个关键的障碍，包括高成本，长时间，以及缺乏跨遥远物种的许多减数分裂特异性蛋白质的序列保守性。为了克服这些障碍，我们已经开发出具有成本效益的基因组学和蛋白质组学方法，系统地确定了大量的未知哺乳动物减数分裂因子。目前的申请将利用遗传学，细胞生物学和生物化学方法的创新组合来确定我们新发现的新的/未表征的因子及其相互作用蛋白在调节小鼠减数分裂重组中的作用。拟议的研究将集中在机制的见解减数分裂重组的调节在三个很大程度上未知或知之甚少的方面：第二端捕获，交叉形成，同源染色体分离。通过减数分裂的进展是生殖细胞体外衍生的关键障碍，并且用于多细胞生物体中体外减数分裂的稳健细胞培养系统仍然难以捉摸。因此，使用一个独特的GFP报告，以监测体外减数分裂的进展，我们计划询问培养参数，以逐步的方式开发一个基于细胞培养的体外减数分裂系统。毫无疑问，哺乳动物减数分裂体外系统的开发不仅将推动减数分裂的基础机制研究，而且将彻底改变人类男性不育症治疗的再生医学。

 DESCRIPTION (provided by applicant): The long-term goal of the current application is to elucidate novel molecular networks underlying mammalian meiosis and develop a mammalian cell culture system for in vitro meiosis. Meiosis, a process unique to germ cells, involves pairing synapsis, recombination, desynapsis, and segregation of homologous chromosomes. Genetic abnormalities resulting from meiosis are a leading cause of birth defects, pregnancy loss, and infertility in humans. The structural and functional properties of meiotic chromatin, which undergoes extensive reorganization, are undoubtedly the central themes of meiosis. Despite rapid progress in understanding meiosis, the complex interplay between chromatin organization and meiotic processes remains largely unexplored. In particular, the progress in understanding mammalian meiosis has lagged far behind meiotic studies in other model organisms, due to several critical barriers, including high cost, long duration, and the lack of sequence conservation of many meiosis-specific proteins across distant species. To overcome these roadblocks, we have developed cost-effective genomics and proteomics approaches to systematically identify a large number of uncharacterized mammalian meiosis factors. The current application will utilize an innovative combination of genetic, cell biological, and biochemical approaches to determine the role of our newly identified novel/uncharacterized factors and their interacting proteins in regulating meiotic recombination in mice. The proposed studies will focus on mechanistic insights into the regulation of meiotic recombination in three largely unknown or poorly understood aspects: second end capture, crossover formation, and homologous chromosome segregation. Progression through meiosis is a critical barrier to in vitro derivation of germ cells and a robust cell culture system for in vitro meiosis in multicellulr organisms remains elusive. Therefore, using a unique GFP reporter to monitor progression of meiosis in vitro, we plan to interrogate culture parameters in a stepwise manner to develop a cell culture-based in vitro meiosis system. Undoubtedly, development of an in vitro system for mammalian meiosis will not only drive basic mechanistic research on meiosis but also revolutionize regenerative medicine in the treatment of male infertility in humans.