TRPM及TRPV家族mRNA在大鼠生精细胞中的表达  

Expression of TRPM and TRPV channel family mRNA in rat spermatogenic cells

作　　者：李世林[1,2] 王行环[2] 王怀鹏[2] 杨中华[2] 高炜城[2] 蒲小勇[2] 

机构地区：[1]南方医科大学研究生学院,广东广州510515 [2]广东省人民医院泌尿外科,广东广州510080

出　　处：《南方医科大学学报》2008年第12期2150-2153,共4页Journal of Southern Medical University

基　　金：广东省自然科学基金(07001203;07001197);广东省医学科研基金(A2007028);中国博士后科研基金(20060400779)

摘　　要：目的探讨瞬时受体电位M型(TRPM)及V型(TRPV)家族基因在大鼠生精细胞中的表达。方法采用机械法分离大鼠生精细胞,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,实时荧光定量RT-PCR技术检测目的基因相对表达量。结果在大鼠生精细胞中检测到TRPM3、TRPM4、TRPM7及TRPV5mRNA的表达,未能检测到TRPM1、TRPM2、TRPM5、TRPM6、TRPM8、TRPV1、TRPV2、TRPV3、TRPV4及TRPV6mRNA的表达。实时荧光定量RT-PCR技术检测显示TRPM家族在大鼠睾丸生精细胞中的表达以TRPM7表达量最高,相当于内参β-actin的(0.0430±0.0034)%,TRPM3和TRPM4表达相对较弱,分别相当于TRPM756%和63%。大鼠睾丸生精细胞中TRPV家族仅存在TRPV5的表达,其表达量相当于内参β-actin的(0.0157±0.0029)%。结论大鼠生精细胞中存在多种TRPM及TRPV家族基因的表达,其中TRPM4及TRPM7mRNA在大鼠生精细胞中呈相对较高表达。本研究结果将为研究生精细胞中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 transient receptor potential melastatin （TRPM） and transient receptor potential vaniUoid （TRPV） channel family genes in rat spermatogenic cells. Methods Rat spermatogenic cells were isolated by a mechanical procedure and the total RNA was extracted using TRIzol reagent. TRPM and TRPV channel family genes were amplified by RT-PCR and the presence of the target genes was detected by agarose gel electrophoresis. The relative gene expression levels were measured by real-time quantitative RT-PCR. Results TRPV5, TRPM3, TRPM4 and TRPM7 mRNAs were expressed in rat spermatogenic cells, hut TRPV1, TRPV2, TRPV3, TRPV4, TRPV6, TRPM1, TRPM2, TRPM5, TRPM6, TRPM7 and TRPM8 mRNAs were not detected. The relative expressions of TRPM and TRPV mRNA were determined by quantitative real-time RT-PCR. TRPM7 expression was the highest among all the TRPM subtypes in rat spermatogenic cells, at a level equivalent to （0.0430±0.0034）% of β-actin expression. TRPM3 and TRPM4 were also highly expressed, but their expression levels were only approximately 56% and 63% of that of TRPM7, respectively. For the TRPV subfamily, only TRPV5 mRNA was abundantly expressed at the level of （0.0157±0.0029）% relative to that of β-actin. Conclusions TRPV5, TRPM3, TRPM4 and TRPM7 mRNAs were coexpressed in spermatogenic cells in rats, among which TRPM4 and TRPM7 mRNA were expressed at high levels. TRPM4 and TRPM7 channels may be involved in the regulation of growth, differentiatiofi and maturation of rat spermatogenic cells and are associated with the generation of the sperms.

关 键 词：瞬时受体电位M型 瞬时受体电位V型 生精细胞 

分 类 号：R346[医药卫生—基础医学]