雄性生殖细胞特异性表达绿色荧光蛋白小鼠的繁殖及其表型鉴定

Production and phenotype identification of specific expressed green fluorescent protein in male mice germ cells

目的繁殖雄性生殖细胞特异性表达绿色荧光蛋白(green fluorescent protein,GFP)小鼠,为进行小鼠生殖系统毒性研究提供有效的工具。方法将Ddx4-cre转基因雄鼠与Rosa26mT/mG转基因雌鼠交配,产生子代动物,利用分子生物学、组织病理学及活体成像等技术,分别从分子、细胞和组织水平对子代及其亲代小鼠进行表型鉴定。结果 PCR结果表明在子代小鼠的睾丸组织中发生了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。

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 Rosa26 mT/m G 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/lox P recombination system were built successfully.