核糖核酸酶H1调节飞蝗嗅觉行为的分子机制研究

核糖核酸酶H1 (RNase H1)作为核酸水解酶，调节体内RNA/DNA杂交体水平，在包括嗅觉以及神经系统在内的多个组织中广泛存在，并和神经退行性疾病有关。但是RNase H1调控嗅觉行为的功能和机制不清楚。本项目以飞蝗为研究模型，通过定量PCR、荧光原位杂交和免疫组化等技术确定RNase H1在脑中分布以及与嗅觉行为的关系，并结合行为以及电生理分析确定RNase H1对飞蝗嗅觉行为调节的基础上，验证RNase H1对RNA/DNA杂交体水平的调节。通过免疫共沉淀测序、转录组测序以及生物信息学分析受RNase H1影响并且形成RNA/DNA杂交体的差异基因。通过生物信息学分析、定量PCR、免疫共沉淀以及嗅觉行为测试等方式分析候选基因是否调节嗅觉行为，其表达是否受RNA/DNA杂交体的调节，进一步研究RNase H1通过调控RNA/DNA杂交体-靶基因环路对飞蝗嗅觉行为调控的神经分子机制。

Ribonuclease H1 locates universally in nervous systems in vertebrates and invertebrates. This enzyme resolves the level of RNA/DNA hybrids and shows close relationship with various neurodegenerative diseases. However, its functions in modulating behaviors, especially olfactory behaviors, in central nervous system are largely unknown. Here we use the migratory locust as an animal model to study the tissue location and the function of RNase H1 in the central nervous system by quantitative PCR, in situ hybridization and immunohistochemistry methods. After confirming the role of RNase H1 in modulating olfactory behaviors by behavior, RNAi knockdown and electrophysiology assays, we further explore its functional relationship with the modulation of the RNA/DNA hybrid level. After then, we apply immunoprecipitation and high throughput sequencing to analyze the expression of the genes forming RNA/DNA hybrids and affecting olfactory behaviors. Moreover, we apply computational analysis, quantitative PCR, immunoprecipitation and olfactory behavioral assay to detect whether the potential genes mediate olfactory behaviors, and their expression levels are affected by RNA/DNA hybrid levels. In this study, we hope to confirm the RNaseH1-RNA/DNA hybrid-targets circuit to mediate neurophysiological mechanism underlying olfactory behavior.