鲟鱼生殖细胞移植研究

项目名称： 鲟鱼生殖细胞移植研究

项目编号： No.31472286

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 李创举

作者单位： 中国水产科学研究院长江水产研究所

项目金额： 88万元

中文摘要： 中华鲟为大型江海洄游性鱼类，由于人类活动的影响，其生存环境恶化，物种处于濒危状态。中华鲟全人工繁殖已获成功，但人工养殖中华鲟性成熟晚且性腺很难发育成熟，迫切需要引进新方法来缩短其繁殖周期。鱼类生殖细胞移植研究进展很快，已在虹鳟、罗非鱼和银汉鱼中获得成功。达氏鲟性成熟时间短，人工繁殖技术成熟，是中华鲟生殖细胞移植的合适受体。本研究拟克隆达氏鲟vasa、dnd和nanos等生殖细胞发育调控基因，研究其在胚胎发育过程中的时空表达特征；进行基因敲降和拯救实验，确定其在达氏鲟生殖细胞发育中的功能；敲降上述所得关键调控基因，制备生殖细胞缺失的达氏鲟作为受体。随后，分离中华鲟精原细胞并移植到不育受体中获得生殖细胞嵌合体；对供体在受体内的发育状况进行监测并采用转录组分析的方法探讨供体生殖细胞在受体内的发育机制。本研究将建立鲟鱼生殖细胞移植技术体系，为中华鲟等濒危动物的物种保护及其资源的可持续利用提供基础。

中文关键词： 中华鲟；达氏鲟；生殖细胞移植；基因功能；转录组

英文摘要： The Chinese sturgeon (Acipenser sinensis) is a large anadromous bony fish, which is currently endangered due to the deteriorated natural habitat caused by anthropogenic activities. Although the artificial propagation of the Chinese sturgeon has already been successfully conducted, many problems exist, such as the rather tough and long period of sexual maturation of the breeding sturgeons. Therefore new methods are badly needed for shortering their reproduction period.. Spermatogonial stem cell transplantation has already been applied to many fishes, such as zebrafish, rainbow trout, Nile tilapia and pejerrey. Considering its mature artificial propagation technique and short period of first sexual maturation, Siberian sturgeon is the appropriate recipient of germ cell transplantation. In the present study, the primordial gonad cell\development-related genes such as dnd, vasa and nanos will be cloned, and the temporal and spatial expression patterns of the acquired genes during embryo development will be studied. In addition, gene specific Morpholino (MO) knockdown and mRNA rescue experiments will be used to explore their biological functions. Finally, MO of suitable germ cell genes will be injected into fertilized eggs of Siberian sturgeon to get the sterile recipients. Mosaic individuals will be obtained by tranplatation of Chinese sturgeon spermatogonial stem cells into the sterile Siberian sturgeon recipients. Bioinformatic analysis like transcriptome sequencing will be adopted for further investigating the developing mechanism of the donor-derived stem cell in the allogeneic recipient. Through this study, spermatogonial stem cell transplantation technique in sturgeons will be established, which should provide the basis for species conservation and sustained resource exploitation of Acipenser sinensis.

英文关键词： Chinese sturgeon;Dabry's sturgeon;germ cell transplant;gene function;transcriptome