Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia  

作　　者：Xin Guo Yao-Ting Gui Ai-Fa Tang Li-Hua Lu Xin Gao Zhi-Ming Cai 

机构地区：[1]The Laboratory of Male Reproductive Medicine, Peking University Shenzhen Hospital, Shenzhen 518036, China [2]Department of Urosurgery, The Third Affiliated Hospital, Sun Yet-Sen University, Guangzhou 510630, China

出　　处：《Asian Journal of Andrology》2007年第3期339-344,共6页亚洲男性学杂志（英文版）

基　　金：We would like to thank Mr Jian-Rong Zhang, Mr Li-Bing Zhang and Dr Zhen-Dong Yu for technical assistance. This work was supported by grants from the National Natural Science Foundation of China （No. 30500543）, Ministry of Education ＂985 project＂ （No. 985-2-054-29）, and Shenzhen Foundation of Science ＆ Technology （JH200505270413B）.

摘　　要：Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.

关 键 词：VASA ejaculated spermatozoa OLIGOZOOSPERMIA male infertility SPERMATOGENESIS 

分 类 号：R698.2[医药卫生—泌尿科学]