基于大鲵重组激活基因、T细胞受体基因和CD基因的时空表达对T淋巴细胞发育时间节点的研究

The Chinese giant salamander (Andrias davidianus) is a unique amphibian and economic species in China. But the development biology of its immune system is obscure. We will focus on the development time points of T lymphocyte in Chinese giant salamander immune system in this study. In our previous studies, we have cloned the marker genes of T lymphocyte such as Rags (recombination activating genes)、TCR (T cell receptor gene) and CD genes then verified the conservative of genes function, which provide the preliminary determination of the development time points of T lymphocyte. In this study, the temporal and spatial expression pattern of the molecular markers in thymus and peripheral blood will be investigated through the technology such as in situ hybridization, Western Blot, immunohistochemistry and realtime PCR to monitor the development time points and location of T lymphocyte. Detection of immune reaction response to antigen of T lymphocyte will be used to verify the development time points. Morphology study of thymus through tissue section and transmission electron microscope techniques will be used to identify the histomorphology and ultrastructural changes of thymus which serve as structural basis of T lymphocyte development. This study would provide invaluable insights into the development of the Chinese giant salamander T lymphocyte, and also provide guideline for resource protection and breeding sustainable development of the Chinese giant salamander.

中国大鲵是我国特有的珍稀两栖动物和重要经济品种，但对其免疫系统发育生物学研究十分薄弱。本项目拟设计实验对大鲵免疫系统中T淋巴细胞的发育时间节点开展研究。我们已克隆T淋巴细胞标记基因(重组激活基因Rags、T细胞受体基因TCR和CD基因) 并验证了其功能的保守性，为T淋巴细胞发育时间节点的确定提供了初步界定。本项目拟采用分子标志物显示技术如原位杂交、免疫组织化学染色、Western Blot 、realtime PCR分析大鲵胸腺和外周血中Rags、TCR和CD基因的时空表达模式，揭示T淋巴细胞的发育时间节点及空间定位；检测T淋巴细胞对抗原的应答反应，进一步验证T淋巴细胞发育的时间节点；同时利用组织切片和透射电镜技术对胸腺各时间节点的组织形态和超微结构变化进行描述，揭示T淋巴细胞发育场所的结构基础。该研究有助于认识大鲵T淋巴细胞发育过程，对大鲵的资源保护和养殖业的可持续发展具有重要指导意义。

中国大鲵是我国特有的珍稀两栖动物和重要经济品种，但对其免疫系统发育生物学研究十分薄弱。本项目对大鲵免疫系统中T淋巴细胞的发育时间节点和发育场所的结构基础开展研究。我们采用分子标志物显示技术如原位杂交、免疫组织化学染色、Western Blot 、realtime PCR分析大鲵胸腺和外周血中Rags、TCR和CD基因的时空表达模式，揭示了T淋巴细胞的发育时间节点及空间定位；检测T淋巴细胞对抗原的应答反应，进一步验证T淋巴细胞发育的时间节点；同时利用组织切片对胸腺各时间节点的组织形态变化进行描述，揭示T淋巴细胞发育场所的结构基础。该研究有助于认识大鲵T淋巴细胞发育过程，对大鲵的资源保护和养殖业的可持续发展具有重要指导意义。