Ropporin基因在人精子中的表达和意义  

The expression and significance of Ropporin in human ejaculated spermatozoa

作　　者：陈静[1] 王勇[1,2] 来永庆[1] 李慧[1] 颜秋霞[1] 桂耀庭[1] 蔡志明[1] 

机构地区：[1]北京大学深圳医院男性生殖与遗传广东省重点实验室,深圳518036 [2]汕头大学医学院生理系

出　　处：《中国男科学杂志》2009年第6期14-18,共5页Chinese Journal of Andrology

基　　金：国家自然科学基金(30770810);广东省科技计划项目(2007B060401061);深圳市科技计划项目(200602059)

摘　　要：目的检测ropporin基因在人精子中的表达。探讨Ropporin抗体对人精子运动的影响,评价Ropporin蛋白在精子运动中的功能。方法收集正常男性精子,非连续梯度离心法分离精子,用RT-PCR、免疫印迹和免疫荧光方法检测ropporin基因在精子中的表达和蛋白定位;将纯化精子分别在含不同浓度Ropporin抗体的培养液中孵育(0,0.8,4和20μg/mL),在不同时间点(1,2,6h)用精子计算机辅助分析系统(computer-assisted sperm analysis,CASA)检测精子活动力。结果RT-PCR、免疫印迹和免疫荧光方法均检测出ropporin基因在精子中表达;免疫荧光表明Ropporin主要定位在精子鞭毛的主段和末段;抗体实验显示抗体孵育1h、2h、6h后精子的快速前向运动(a级)和慢速前向运动(b级)均明显受到抑制,并显示剂量依赖性。结论Ropporin在精子鞭毛的主段和末段上的表达与精子的运动密切相关。Objective To detect the expression and distribution of Rropporin in human spermatozoa, and evaulate the function of Ropporin in human sperm motility. Methods Human sperm from normozoospermic donors were collected for analysis of ropporin expression The mRNA and protein expression of Ropporin were detected reverse transcription polymerase chain reaction （RT-PCR）, western blot and immunoflorescence, respectively. The isolated spermatozoa from normozoospermicdonors by Percoll discontinuous density gradients were then incubated with different ropporin antibody at final concentrations of 0, 018, 4and 20μg/mL. After 1, 2 and 6 h incubation, sperm motility grades including progressive motility, fast progressive motility and slow progressive motility were measured by means of computer-assisted sperm analysis. Results The expression of Ropporin transcript was all detected in each human semen sample. Distribution of Ropporin protein was localized at the principle segment and end segment of sperm flagella. Fast progressive motility and slow progressive motility of sperm were all inhibited under the use of Ropporin antibody in different concentrations and-moreover showed a significant dose-dependent changes were observed. Conclusion Distribution of Ropporin at the principle segment and end segment of sperm flagella might be closely associated with sperm motility.

关 键 词：ropporin基因 基因表达 精子能动性 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]