精子液态储存的分子机制-基于中华鳖输卵管模式

精子液态储存是某些动物进化的一种特殊生殖策略，能有效延长受精时间，提高繁殖成功率。课题组前期的实验发现，中华鳖液态精子储存现象典型，储存部位的上皮与精子之间联系紧密，且上皮细胞的雄激素受体呈阳性免疫反应。本项目以此为切入点，将在细胞和分子水平上，揭示精子液态储存期雄激素水平的变化规律；显示氟他胺（雄激素拮抗剂）处理前后，储存精子的结构和活力的变化特征，定量分析雄激素受体、Bcl-2、Caspase-3和MAP激酶在输卵管峡部和子宫部（精子储存微环境）的表达规律；进一步通过输卵管上皮组织的体外培养，检测睾酮或睾酮与氟他胺联合作用后，上述相关蛋白及其基因的表达动态，进而分析雄激素依赖性信号途径在峡部和子宫部的调节作用。通过体内和体外实验，从不同角度阐明雄激素及其调控系统在中华鳖输卵管（天然模型）精子液态储存中的作用机理。研究结果可为体外精子液态保存新技术的建立提供独特思路和理论支撑。

Sperm storage in liquid is a special reproductive strategy evolved by some animals, which can extend the period of fertiliation effectively, and increase reproductive ratio successfully. Our previous work has revealed that Chinese soft-shelled turtle can be a typical model in long-term sperm storage. Morphological relationship between sperm and epithelium is very close, and the immune reaction of androgen receptor in storage region is positive. This project will show the diversify pattern of androgen level during sperm storage by cellular and molecular methods. Structure and mobility of stored sperm are examined before and after flutamide (anti-androgen) treatment. Expression of androgen receptor,Bcl-2, Caspase-3 and MAP kinase in isthmus and uterine part of oviduct is analized quantitatively. Proteins above and their genes on the epithelium of cultured oviduc are further investigated after treatment by testosterone or testosteroneand flutamide in vitro. Regulation of androgen and its signal pathway in liquid sperm storage of soft-shelled turtle will be elucidated according the reseach experiments in vivo and in vitro. Results of this project could provide an theoretical enlightenment for the development of new technology in liquid sperm conservation in vitro.