精原干细胞中Pten基因敲除小鼠模型的建立及初步的表型分析  

Establishment and Preliminary Phenotypic Analysis of Pten Knockout Mouse Model in Spermatogonial Stem Cells

作　　者：周炜[1] 张迪[1] 于卓[1] 查巍巍[2] 冯立新[1] 

机构地区：[1]上海交通大学医学院,上海200025 [2]中国林业科学研究院,北京100091

出　　处：《实验动物与比较医学》2014年第1期1-5,共5页Laboratory Animal and Comparative Medicine

基　　金：国家自然科学基金（31171409）

摘　　要：目的 建立稳定的精原干细胞中Pten基因敲除的小鼠模型,对表型进行初步观察,为研究Pten在精原干细胞中的作用及相关的机制奠定基础.方法 采用Cre/Loxp系统建立Pten在精原干细胞中条件性敲除的小鼠模型,以同窝正常雄鼠作为对照,通过解剖、HE染色等方法对小鼠的表型进行观察,并通过免疫荧光检测精原干细胞标记分子早幼粒细胞白血病锌指蛋白（Plzf）的表达并对免疫荧光染色中的阳性细胞进行统计.结果 与对照组相比,Pten敲除鼠在出生后出现发育迟缓现象,13d左右个体死亡,睾丸大小未受影响,但进一步的细胞计数统计发现Pten敲除后Plzf阳性细胞数目和对照组相比明显减少（P＜0.01）.结论 成功建立精原干细胞中Pten基因敲除的小鼠模型,并且Pten可能通过下调Plzf的表达影响精原干细胞的发育,这为进一步探讨精原干细胞复杂的调控网络提供了一定的实验依据和思路.Objective To establish Pten conditional knockout （KO） mouse model and investigate the role and related mechanism of Pten in spermatogonial stem cells（SSCs） and preliminary animal phenotype. Methods Cre/Loxp system was adopted to establish the Pten conditional knockout（KO） mouse model in spermatogonial stem cells. The wild type littermates were used as control and animal phenotype was observed by dissection and Hematoxylin and Eosin （HE） Staining. Then the expression of Plzf, a kind of SSCs marker, was detected by immunofluorescence stain （IF） and the positive cells in IF were counted. Results Pten KO mice shown postnatal developmental retardation and some died around 13 days. No difference in the size of testes was found comparing to the wild type mice, but the result of cell counting indicated that less Plzf positive cells were detected in KO mice （P〈0.01）. Conclusion Pten conditional knockout mouse model using spermatogonia stem cells is successfully established and Pten might affect SSCs development by regulating the expression of Plzf. This study, to some extent, provides some scientific evidence and new thoughtfor further exploring the complicated regulatory network in SSCs.

关 键 词：CRE loxp系统 Pten基因敲除 精原干细胞 表型分析 锌指蛋白(Plzf) 

分 类 号：Q95-33[生物学—动物学]