细胞色素蛋白Cyt-c1L在果蝇精子个体化过程中的功能及作用途径

精子发生及基因调控机制是发育生物学研究的热点之一。果蝇精子个体化过程是产生有功能的成熟精子的必要条件，这一过程涉及到线粒体结构与功能的变化。Cyt-c1L在线粒体电子传递过程中发挥作用，在雄蝇（精巢）中特异性表达，但其具体功能还不清楚。我们前期发现在黑腹果蝇ocnus(ocn)敲降精巢中，Cyt-c1L表达显著下调，Cyt-c1L敲降使精子个体化复合物不能形成，成熟精子消失，导致雄性不育。本项目拟先通过ChIP-qPCR、EMSA研究Ocn对Cyt-c1L的调控作用；再通过基因敲除/敲降、免疫荧光等方法探讨Cyt-c1L对线粒体信号传导以及Caspase途径的影响机制；然后通过研究精细胞延长中动力蛋白的动态，探究Cyt-c1L影响果蝇精子个体化的信号通路。预期结果既可以丰富精子成熟过程中基因调控机制的基础理论，也可为人类及其他动物的雄性不育和线粒体疾病研究提供理论参考。

Spermatogenesis and gene regulation mechanisms are one of the hotspots in developmental biology research. The sperm individualization process of Drosophila is necessary for the production of functional mature sperm, in which the mitochondrial structure and function of are involved. Cyt-c1L plays a role in mitochondrial electron transport and is specifically expressed in male fruit flies, especially in testes, but its function are not fully understood. In previous, we found that the germline stem cells are missing in the ocnus (ocn) knockdown testis. Cyt-c1L expression is significantly down-regulated when ocn was knock down in testes. Cyt-c1L knockdown can cause sperm individualized complex (IC) fail to form properly, inducing that mature sperm in the seminal vesicle are absent, thus causing male sterility. In this research, we intend to investigate the regulation of Cyt-c1L by Ocn through ChIP-qPCR and EMSA method, and then explore Cyt-c1L knock down effect on mitochondrial signaling and caspase activity in testes through gene Knock out/down, immunofluorescence. In addition, we study the dynamic protein changes to explore the signal pathway of Cyt-c1L affecting the sperm individualization. The expected results can not only enrich the basic theory of gene regulation mechanism during sperm maturation, but also provide a theoretical reference for mechanisms of study on male sterility and mitochondrial disease in humans and other animals.