热休克蛋白HSP70活化特征在轮虫衰老过程中的作用及其调控分子机理研究

Rotifera, which distributed widely,easy to culture, smaller(<0.5mm) and reproductive speed quickly (offsprings were produced interval 2 hour after they matures physically ), and has a shorter life span in fresh water body, It is a new field for rotiferological research that the activation feature of heat shock protein 70 and the mechanism of modulation of JNK and TOR pathway in aging process of rotifer.. In this project, Brachionus calyciflorus widely distribute in the world was selected as test animal by using their neonate hatched from resting eggs as initial material. A steady clone will be established by using an amictic female initial hatched from resting eggs of rotifer..Rotifers at different age were undergo stress (heat shock, oxidation material, UV-B radiation and starvation). Full cohort life table will be used to analyze characteristic of rotifer aging population. The feature of heat shock protein 70 activated in aging should be investigated by using methods of RNA interference, gene knockout，Immunohistochemistry and Western blotting and then to reveal the molecular mechanism of Hsp70 modulate JNK and TOR pathway in rotifer aging and then promote the research of aging in rotifer.

轮虫分布广、易于培养和个体微小(<0.5mm)、生殖速度快(性成熟后，每隔2h产生1次后代)、生命周期短(10d左右)，是研究衰老生物学的新型受试动物。热休克蛋白HSP70在轮虫衰老过程的活化特征、JNK和TOR信号通路在调控轮虫衰老过程中的作用,是近年来轮虫学研究的热点领域。本项目采用淡水广布种萼花臂尾轮虫为受试动物，利用单个母体进行“克隆”群培养，建立实验品系。采用休眠卵孵化的幼体（日龄<2h），进行胁迫刺激(高温刺激、过氧化物胁迫、UV-B辐射和饥饿等)。经历胁迫后，分析轮虫1、3、5、7和10日龄热休克蛋白HSP70、JNK和TOR的活性特征。利用RNA干扰、基因敲除、免疫组化和Western blotting等技术方法，研究轮虫衰老过程的Hsp70蛋白活化特征；HSP70对JNK和TOR信号通路的调控作用；三者在轮虫衰老中的作用途径及其分子调控机理等，推动轮虫衰老生物学研究。

轮虫是目前研究衰老生物学的新型潜在受试动物之一，热休克蛋白HSP70在轮虫衰老过程的活化特征、JNK和TOR信号通路在调控轮虫衰老过程中的作是近年来轮虫学研究的热点领域。本项目采用轮虫为受试动物，研究了槲皮素、原花青素、黄连素和二甲双胍等不同外源性物质对轮虫生殖（生殖前期、生殖期）、寿命和生理生化指标等特性的影响。利用实验生态学和分子生物学技术比较研究了轮虫衰老过程中hsp40、hsp70和hsp90在轮虫衰老过程中变化规律和活化特征、雷帕霉素对TOR的影响，对不同剂量和不同作用时间下轮虫体内TOR动态特征及表达规律进行了系统研究。根据轮虫不同发育阶段HSP70的变化，通过抑制或干扰JNK比较研究了hsp70、JNK在轮虫衰老过程中的动态变化关系和彼此作用的调控作用，揭示了热休克蛋白尤其是hsp70在轮虫衰老中的作用及其分子调控机理，推动了轮虫衰老生物学研究。