Plac8l1在小鼠精子发生过程中的表达特征研究  

Expression characteristics of Plac8l1 in mouse testicular tissue

作　　者：王慧英[1] 张蕊[1] 张瑾[1] 昌文林 Wang Huiyin;Zhang Rui;Zhang Jin;Chang Wenlin(Department of Obstetrics and Gynecology,Beijing Shijitan Hospital, Capital Medical University;Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics ,Institute of Urology, Peking University Shenzhen Hospital,Biomedical Research Institute ,Shenzhen Peking University-The Hong Kong University of Science and Technology Medical Center)

机构地区：[1]首都医科大学附属北京世纪坛医院妇产科,北京100038 [2]男性生殖与遗传广东省和深圳市重点实验室北京大学深圳医院泌尿外科研究所深圳北京大学香港科技大学医学中心,深圳518036

出　　处：《重庆医科大学学报》2018年第3期410-413,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:31701305);深圳市科创委2017年度基础研究(自由探索)资助项目(编号:JCYJ20170306161657672)

摘　　要：目的:探讨Plac8l1(placenta-specific 8 like 1)基因在小鼠睾丸组织的表达特征,及其与精子发生的相关性。方法:经生物信息学方法初步筛选到一个睾丸特异性表达候选基因Plac8l1,并推测其可能与小鼠精子发生相关;采用半定量RT-PCR和实时定量RT-PCR对Plac8l1表达的时空特异进行了研究;应用原位杂交技术对Plac8l1 m RNA在成年小鼠睾丸中的细胞类群表达特征进行初步研究;通过构建过表达PLAC8L1-EGFP重组蛋白质粒及生物信息学分析进行亚细胞定位研究与功能预测。结果:(1)Plac8l1是1个睾丸特异性转录基因。(2)在成年小鼠,Plac8l1只在睾丸和附睾组织发生特异性转录,而且睾丸中的表达水平明显高于附睾[睾丸vs.附睾=[(1.000±0.000)vs.(0.036±0.018),P=0.000]。(3)随着小鼠精子发生的起始,睾丸中Plac8l1转录水平逐渐升高(P=0.000)。(4)m RNA原位杂交结果显示Plac8l1 m RNA主要定位在成年小鼠睾丸中的间质细胞和精母细胞。(5)经生物信息学分析,PLAC8L1也有着一个类似于PLAC8的功能域,过表达重组蛋白显示PLAC8L1在细胞膜上聚集。结论:Plac8l1是1个睾丸特异性基因,可能与精子发生过程中的精母细胞分化相关;PLAC8L1具有1个富含半胱氨酸的PLAC8结构域,这一结构域可能介导了PLAC8L1在细胞膜上聚集并与其他蛋白相互作用,并可能在精母细胞分化中发挥重要作用。Objective ：To study the expression characteristics of Placenta-specific 8 like 1 （PlacSll） in mouse testicular tissue,and to reveal its possible function during spermatogenesis. Methods：The candidate gene PlacSll was preliminary screened to be specific ex- pression in testis by bioinformatics methods,and it is speculated that the gene was related to the spermatogenesis. Besides,semi quantitative RT-PCR and real-time quantitative RT-PCR were used to investigate the tissue specificity of placSl and the temporal and spatial characteristics of the process of spermatogenesis,while in situ hybridization was used to preliminarily study the expression characteristics of PlacSll mRNA in the testis of adult mouse. Finally,the subcellular localization was studied and its functional was predicted by constructing PLAC8L1-EGFP recombinant protein particle and bioinformatics analysis. Results：（1）The results of preliminary bioinformatics showed that Plac811 is one of gene for specific transcription. （2）Semi-quantitative and quantitative real-time RT-PCR indicated that Plac8ll only transcript in adult mice testis and epididymis,and testieular expression level was significantly higher than epididymis. （3）As the mouse matured, the PlacSll transcription level in testis increased gradually. （4）RNA in situ hybridization showed that mRNA was mainly positioned in ledig cell and sperm mother cell in mature mouse testis. （5）Bioinformatics analysis showed that PLAC8L1 also had a function domain similar to the PLAC8 and the over expression of recombinant proteins showed that protein aggregated in the cell membrane. Conclusion：Plac811 is a testis specific gene,and is very likely to associate with spermatocyte differentiation in the process of spermatogenesis. PLAC8L1 has a cystine which riches in PLAC8 domain,and the domain may mediate PLAC8L1 to aggregate in cell membrane and interact with other protein and play an important role in spermatocyte differentiation.

关 键 词：睾丸特异性基因 Plac8l1 精子发生 睾丸 小鼠 

分 类 号：R318.16[医药卫生—生物医学工程]