孕酮激素诱发斑马鱼精原细胞增殖分化的机理研究

项目名称： 孕酮激素诱发斑马鱼精原细胞增殖分化的机理研究

项目编号： No.31201977

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 陈仕玺

作者单位： 厦门大学

项目金额： 22万元

中文摘要： 近年来，已报道的和我们前期开展的研究结果初步表明，鱼类特有的孕酮激素- - 17α,20β-双羟孕酮（DHP）参与了精原细胞增殖分化过程的调控。同时，应用斑马鱼开展的研究已经明确，孕酮激素核受体（nPR）在下丘脑视前区以及精巢内多种细胞上均有表达。为了进一步阐明孕酮激素诱发斑马鱼精原细胞增殖分化的机理，本项目拟在前期研究工作的基础上，开展以下研究内容：（1）应用活体暴露的方法，研究DHP是否参与下丘脑-垂体系统中生殖神经内分泌激素的调控；（2）应用以A型精原细胞为主的精巢进行体外培养实验，研究DHP对精原细胞增殖分化的作用；（3）应用荧光激活细胞分选法从转基因斑马鱼精巢中分离出精原细胞和支持细胞,再结合抑制性消减杂交技术和蛋白质组学技术，研究DHP对精原细胞和支持细胞基因表达的调控。研究结果不仅可以丰富鱼类生殖内分泌调控研究的基础理论，也可为今后我国鱼类人工繁殖技术的完善提供科学依据。

中文关键词： 斑马鱼；孕酮核受体；生殖神经内分泌；精巢；

英文摘要： Recently, research reports showed that 17α,20β-dihydroxy-4-pregnen-3-one (DHP), the biologically active progesterone molecule in many teleost fishes, has multiple regulatory functions during the proliferation and differentiation of spermatogonia. Our previous studies showed that, in zebrafish, exposure to DHP in vivo induced the proliferation of type A spermatogonia and their proliferation and differentiation into type B spermatogonia and also spermatocytes. Besides, after characterization of the zebrafish nuclear progestin receptor (nPR), further analyses indicated that the nPR was expressed in spermatogonia and Sertoli cells and Leydig cells in the testis. Moreover, another study indicates nPR was expressed robustly in the preoptic region of the hypothalamus in the brain. The localization of Pgr suggests that it mediates DHP regulation of spermatogonial proliferation and differentiation. The present study aims at elucidating the molecular mechanism(s) of these DHP effects. Using the zebrafish as model animal, we propose to investigate the (1) potential effect of DHP on the regulation of reproductive neuroendocrine hormones in the hypothalamus-pituitary; (2) effects of DHP directly on testis enriched in type A spermatogonia, by using a recent developmental primary testis tissue culture system; (3) the target ge

英文关键词： zebrafish；nuclear progesterone receptor；reproductive neuroendocrinology；testis；