睾丸特异性表达绿色荧光蛋白的转基因小鼠模型的建立.doc

雄性生殖细胞特异性表达GFP小鼠的繁殖及其表型鉴定 王志茹,2，李军2，刘晓梅1，吴红联2，朱德生2 吉林大学公共卫生学院，长春2. 北京大学实验动物中，北京00871） 【摘要】目的 繁殖雄性生殖细胞特异性表达绿色荧光蛋白（小鼠Cre酶介导的特异性基因重组；活体成像可以看到在F1代小鼠的睾丸组织中具有GFP的表达；睾丸冰冻切片及精子荧光观察显示GFP主要表达于次级精母细胞、精子细胞和精子中。结论 雄性生殖细胞特异性表达GFP小鼠繁殖成功。 【关键词】睾丸；特异性；基因重组；GFP Production and?phenotype identification of?specific expressed green fluorescent?protein in male mice germ cells WANG Zhi-ru1,2，LI Jun2，LIU Xiao-mei1，WU Hong-lian2，ZHU De-sheng2 School of public health，Jilin University，Changchun 130021，China；2. Peking University Laboratory animal centre，eijing 100871，China) 【Abstract】 Objective The aim of this study is production of organ specific animal model for studying reproductive toxicity in mice. Methods F1 generation was gotten by mating the Ddx4-cre transgenic male mice with the Rosa26mT/mG transgenic female mice. F1 offspring and its parents phenotype was screened by molecular biological, histopathological and in vivo imaging technology. Results At molecular level, specific DNA fragment was only found in testis of F1 offspring ; At the organ level, the expression of green fluorescent protein could only be observed in testis of F1 offspring; Testicular frozen sections and sperm fluorescence observation showed that green fluorescent protein were mainly expressed in the germ cell lineage such as secondary spermatocyte and spermatocyte and spermatozoon. Conclusions The production of the mice with specific germ cell expressed green fluorescent protein by Cre/loxP recombination system were built successfully. 【Key words】Testis;Specificity; Gene recombination; Green fluorescent?protein 生殖系统的改变是造成不孕不育的重要原因，如何简易快速的检测生殖系统的损伤是目前研究的一个热点。在绝大，Ddx4[1]，常被用作分子标记研究配子发生和原始生殖细胞的起源、迁移、分化等方面。雄性生殖细胞的 DDX4 蛋白表达，从12.5dpc生殖嵴一直持续到减数分裂后的精子细胞[2]。本研究利用生殖系统特异性启动子Ddx4启动的Cre酶，通过Cre/loxP位点特异性重组系统[3]产生雄性生殖细胞特异性表达GFP小鼠模型，为后续进行雄性小鼠生殖系统发育和毒性研究提供理想的动物模型。 材料与方法 实验动物 SPF级B6.FVB-Tg(Ddx4-cre)1Dcas/JnjuDdx4-cre小鼠（以下简称为Ddx4-cre小鼠B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato,-GFP)Luo小鼠（以下简称Rosa26mT/mG小鼠）4只，雌雄各半，均为纯合子（mut/mut