中华鲟为极度濒危鱼类，其全人工繁殖虽已突破，但养殖亲鱼数量有限且初次性成熟时间过长，亟需新途径来加强其种质资源保存及物种保护。精原干细胞具有分化为两性配子的潜能，建立其体外长期培养体系是构建濒危鱼类保护新途径的关键。GDNF/GFRα1在哺乳动物精原干细胞的自我更新和增殖调控中起关键作用。本研究拟鉴定中华鲟gdnf和gfrα1基因并揭示其表达模式；通过在前期建立的中华鲟精原干细胞培养体系中添加GDNF和GFRα1重组蛋白，研究其对精原干细胞的增殖、凋亡及分化等特性的影响，阐明重组蛋白在中华鲟精原干细胞体外增殖的作用；进一步对重组蛋白添加前后培养的精原干细胞进行转录组测序，比较分析相关信号通路并进行功能验证，探究GDNF/GFRα1调控中华鲟精原干细胞体外增殖的作用机制。本研究的结果将为中华鲟精原干细胞体外长期培养体系的建立提供有效途径，进而为中华鲟的种质资源保存及物种保护提供新的技术支撑。

The Chinese sturgeon (Acipenser sinensis) is a critically endangered species. Although complete artificial propagation of Chinese sturgeon was achieved, only a few brood fish is capable of spawning, and Chinese sturgeon is an extremely late maturing species. Thus, it is urgent to apply new technology in preserving germplasm resources and species conservation of Chinese sturgeon. Spermatogonial stem cells (SSCs) have the ability to differentiate into sperm and eggs, and establishment of its long-term culture system in vitro will be highly significant for the conservation of endangered species. The GDNF/GFRα1 signal pathway plays a critical role in the SSCs self-renewal and proliferation in mammals. In the present study, we will report the identification of gdnf and gfrα1 genes, and their expression pattern at both RNA and protein levels in the Chinese sturgeon. In order to determine the function of GDNF and GFRα1 recombinant proteins in the proliferation of Chinese sturgeon SSCs in vitro, the GDNF and GFRα1 will be added into the SSCs culture system established previously, and then the proliferation, apoptosis and differentiation of cultured Chinese sturgeon SSCs will be analyzed. Furthermore, by means of transcriptome sequencing of cultured Chinese sturgeon SSCs before and after the GDNF and GFRα1 added, the signal pathways related to the SSCs proliferation will be filtered and confirmed in vitro, which will be beneficial to elucidate the mechanism of GDNF/GFRα1 in regulating the proliferation of Chinese sturgeon SSCs in vitro. In a word, this study will provide a possibly effective means for establishing a long-term culture system for the Chinese sturgeon SSCs, which will provide new technological support for the germplasm preservation and species conservation of Chinese sturgeon.