高原鼠兔体细胞中精子特异性乳酸脱氢酶基因的表达对其低氧适应能力的作用及生理机制

项目名称： 高原鼠兔体细胞中精子特异性乳酸脱氢酶基因的表达对其低氧适应能力的作用及生理机制

项目编号： No.31260512

项目类型： 地区科学基金项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 魏登邦

作者单位： 青海大学

项目金额： 50万元

中文摘要： 低氧是青藏高原最鲜明的气候特征。高原鼠兔是一种青藏高原特有的啮齿动物，生活在海拔3 000m以上的高寒草原。高原鼠兔在长期进化过程中形成了一系列适应低氧环境的生理机制,对低氧具有很强的耐受性。我们最近的研究结果表明，Ldh-c基因不仅在高原鼠兔睾丸中表达，而且在其体细胞中表达。我们认为高原鼠兔体细胞中Ldh-c基因的表达，提高了组织无氧糖酵解的水平，减小了在低氧环境中对氧的依赖，增强了对低氧环境的适应能力。因此，本研究通过制备LDH-C亚基特异性抗体，应用免疫组织化学方法研究LDH-C4在高原鼠兔体细胞中定位；通过构建高原鼠兔Ldh-a，Ldh-b和Ldh-c基因的原核表达系统，分离纯化LDH-A4，LDH-B4 和LDH-C4，比较研究它们的酶促动力学特征；应用基因沉默技术研究LDH-C4对高原鼠兔抗缺氧和抗疲劳的作用及生理机制，进一步阐明高原鼠兔适应低氧环境的特殊生理机制。

中文关键词： 高原鼠兔；精子特异性乳酸脱氢酶；酶促动力学；特异性抑制剂；RNAi

英文摘要： Hypoxia is the most distinct climate character of Qinghai-Tibet Plateau. Plateau pika (Ochotona curzoniae) is a native rodent that inhabits in grassland at elevation above 3000m on Qinghai-Tibet Plateau.In evolution, plateau pika has obtained a serious mechanisms to adapt the hypoxic environment, and became a hypoxia tolerant mammal. In recent, our studying results indicated that Ldh-c is expressed not only in testis but also in somatic tissues of plateau pika. We suggested that,by the expression of Ldh-c in somatic tissues, plateau pika decreased dependence on oxygen and enhanced adaptation to the hypoxic environment by increaseing anaerobic glycolysis levels in its somatic tissues.Therefore,to further shed light on the specific adapting mechanisms of plateau pika to the hypoxic enviroment, in present project, we project to research the location of LDH-C4 in plateau pika somatic tissues by immunohistochemistry methods, the catalyzing enzymatically characteristcs of LDH-C4 by comparason with that of LDH-A4 and LDH-B4, and the antihypoxic and antifatigue effects of LDH-C4 on plateau pika as well as its physiological mechanisms by gene silencing methods.

英文关键词： Plateau pika (Ochotona Curzniac)；LDH-C4；Enzyme kinetics；N-isopropyl oxamate；RNAi