线粒体是提供细胞能量，维持细胞稳态的重要细胞器。在细胞分裂过程中，线粒体有效分配到子代细胞是维持子代细胞正常功能和生物世代延续的保障。在线虫精子发生过程中，线粒体被完全分配到精子细胞中，而残余体中没有保留。线粒体的不对称分配过程，及其调控机制尚不清楚。我们发展了线虫精母细胞的体外培养体系，采用特异染料和荧光蛋白报告体系标记不同细胞器和细胞骨架，对线虫精子发生过程中细胞质不对称分配过程进行追踪和观察。本课题拟采用这一线虫精母细胞体外培养体系结合多种遗传学和细胞生物学手段，开展遗传突变筛选，分离鉴定调控线粒体不对称分配的基因突变。同时，结合利用RNA干扰技术、遗传突变和多种显微成像技术，分析线粒体不对称分配的动态变化，鉴定分析已知基因突变对这一过程的作用和影响。

Mitochondria are responsible for generating energy, maintaining cellular homeostasis. During cell division, mitochondria, as well many other organelles, need to properly segregate into two daughter cells. In C. elegans spermatogenesis, mitochondria asymmetrically segregate into spermatid, together with nucleus and fibrous membrane organelle FB-MO. Other cytoplasmic components will deposit to residual body, proceeding to phagocytosis. The mechanism controlling mitochondria asymmetric segregation in spermatogenesis is largely unknown. We have established an in vitro culture system for worm spermatocyte, allowing us to observe the whole spermatocyte division process. Using fluorescence dyes and fluorescence reporters, we are able to trace mitochondria, other organelles and cytoskeleton dynamic changes during spermatid separation. Here, we propose to do a forward genetic screen to identify mutants having mitochondria asymmetric segregation defects and map genes affected by these mutants. Analysis of their functions in C. elegans and in vitro culture will provide insights to mechanisms regulating mitochondria segregation during spermatogenesis. We also propose to conduct the candidate gene search using RNAi library and genetic mutants to analyze genes involved in this process.