摘要
本研究旨在探讨Wip1基因的表达及其对精子受精能力的影响,为阐明Wip1基因对雄性繁殖的影响提供新的着眼点。选取相同饲养环境、同一遗传背景的8周龄左右的野生型雄鼠,利用实时荧光定量PCR技术检测Wip1基因在心脏、肝脏、脾脏、肺脏、肾脏、脑组织及雄性生殖器官中的表达情况,利用免疫荧光技术检测Wip1蛋白在睾丸组织及精子细胞中的分布情况。以Wip1敲除型雄鼠为试验组,野生型雄鼠为对照组,利用ELISA试剂盒检测两组小鼠血清中睾酮水平;通过体外受精试验比较两组的2-细胞率及囊胚率。结果显示,Wip1基因mRNA在野生型雄鼠各组织中广泛表达,且在雄性生殖器官中表达量较高,其中睾丸中表达量最高,附睾体、附睾头、附睾尾次之;Wip1蛋白主要定位于睾丸组织中的长形精子细胞及附睾成熟精子细胞的头部。ELISA检测结果发现,与野生型雄鼠相比,Wip1敲除型雄鼠血清内睾酮含量极显著下降(P<0.01)。体外受精结果表明,Wip1敲除后2-细胞率及囊胚率均极显著下降(P<0.01)。结果表明,Wip1基因敲除能够引起雄性小鼠受精能力降低,这可能与Wip1基因在精子发生过程中发挥作用相关。
This study was aimed to investigate the expression of Wip1 gene and its effect on the fertilization ability in mice,in order to provide a new perspective for further study on the function of Wip1 gene on male reproduction.8-week-old wild type mice from the same environment and genetic background were selected,the expression profile of Wip1 in heart,liver,spleen,lung,kidney,brain and male reproductive organs were analyzed by Real-time quantitative PCR.The distribution of Wip1 protein in testes and sperm was detected by immunofluorescence.Wip1 gene knockout male mice were used as experimental group,and wild type male mice were used as control group,the serum level of testosterone was tested by ELISA.The rates of 2-cell and blastocyst were compared between two groups by in vitro fertilization experiment.The results showed that Wip1 gene was widely expressed in various tissues of wild male mice,and which was higher in male reproductive organs,the expression in testis was the highest,followed by corpus epididymidis,caput epididymidis and cauda epididymidis.Wip1 protein mainly located in elongating spermatids of testes and mature sperm head.ELISA results showed that compared with the wild type male mice,the level of testosterone was extremely significantly decreased in Wip1 gene knockout male mice(P<0.01).In vitro fertilization test results showed that the rates of 2-cell and blastocyst were extremely significantly decreased in Wip1 gene knockout male mice comparing with the wild type mice(P<0.01).In summary,this study found that Wip1 gene knockout could reduce the fertilization ability of male mice,which might be related to the role of Wip1 gene in the process of spermatogenesis.
作者
邱乙卿
高倩
魏迎辉
刘莎莎
牟玉莲
李奎
QIU Yiqing;GAO Qian;WEI Yinghui;LIU Shasha;MU Yulian;LI Kui(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China)
出处
《中国畜牧兽医》
CAS
北大核心
2019年第10期3007-3015,共9页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31572378)
中央级公益性科研院所基本科研业务费专项资金项目(2017ywf-zd-9)
中国农业科学院科技创新工程(ASTIP-IAS05)
