TRPV离子通道家族在人精液生殖细胞中的表达及定位  

The Expression of TRPV Channel Families in Germs of Human Ejaculated Semen

作　　者：李世林 王怀鹏[2] 蔡伟山 

机构地区：[1]广东省广州市南沙中心医院外科,广东广州511457 [2]广东省人民医院泌尿外科,广东广州510080

出　　处：《临床医学工程》2011年第7期979-981,984,共4页Clinical Medicine & Engineering

基　　金：广州市医药卫生科技项目基金(№.201102A213085)

摘　　要：目的探讨TRPV离子通道家族在人精液生精细胞及精子中的表达情况及细胞定位。方法采用非连续Percoll密度梯度离心法分离人精液中生精细胞及精子,TRIzol法提取细胞总RNA,RT-PCR合成并扩增目的基因,琼脂糖凝胶电泳检测目的基因的存在。蛋白印迹技术检测TRPV离子通道蛋白在人精液生精细胞及精子中的表达,免疫荧光染色技术检测TRPV离子通道蛋白在人精液生精细胞及精子中的表达定位。结果在人精液生精细胞及精子中检测到TRPV1、TRPV2及TRPV5mRNA的表达,未能检测到TRPV3、TRPV4及TRPV6mRNA的表达。蛋白印迹检测显示人精液生殖细胞中存在TRPV1、TRPV2及TRPV5离子通道蛋白的表达。免疫荧光染色显示,TRPV1离子通道主要表达于中晚期生精细胞胞浆及精子头部。结论人精液生精细胞及精子中存在多种TRPV家族离子通道的表达,其中TRPV1及TRPV2mRNA在人精液生精细胞及精子中呈相对较高表达,其主要定位于中晚期生精细胞胞浆及精子头部。本研究结果将为研究人类生精细胞及精子中TRP通道的功能及TRP通道各亚型之间的相互关系提供参考依据。Objective To investigate the expression of TRPV channel families in germs of human ejaculated semen and to localize the existence of TRPV ion channels in germs of human semen.Methods The germs in human ejaculated semen were isolated by discontinuous Percoll gradient and total RNA was extracted from dissociated human spermatozoa by using TRIzol reagent.TRPV channel families genes were amplified by RT-PCR and the existence of the target gene was detected by agarose gel electrophoresis.The channel proteins in dissociated human germs were checked by western blot analysis.The distribution of channel proteins in dissociated human spermatozoa was detected by immunofluorescence staining.Results TRPV1,TRPV2 and TRPV5 mRNA were expressed in human spermatogenic cells,but TRPV3,TRPV4 and TRPV6 mRNA were not detected in human germs.Western blot analysis indicated the expression of TRPV1,TRPV2 and TRPV5 channel proteins in human spermatozoa.Immunofluorescence staining showed that TRPV1 channel protein was chiefly expressed in the head and midpiece of flagellum of sperm.Conclusions TRPV1,TRPV2 and TRPV5 mRNA are co-expressed in hu man ejac ulated spermatozoa,among which TRPV1 and TRPV2 mRNA are expressed higher than the others.The results of this study will provide reference basis for investigating TRP channels function and the interrelationship of TRP channel subtypes in human spermatogenic cells and sperm.

关 键 词：瞬时感受器电位V型 生殖细胞 精液 精子 

分 类 号：R698.2[医药卫生—泌尿科学]