Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia
Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia
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正常精子男性与少精子症患者射出精子中VASA基因的差异表达
DOI：
10.1111/j.1745-7262.2007.00253.x
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发表时间：
2007-05-01
影响因子：
2.9
通讯作者：
Cai, Zhi-Ming
中科院分区：
文献类型：
--
作者：
Guo, Xin;Gui, Yao-Ting;Cai, Zhi-Ming
目的：目的检测VASA在人类射出精子中的表达，并比较VASA在正常精子和少精子症患者中的表达。研究方法：通过手淫从正常精子症男性和少精子症患者中收集射出的精子，随后通过Percoll不连续梯度分离以获得精子。采用逆转录聚合酶链反应（RT-PCR）、定量RT-PCR（QRT-PCR）、免疫荧光和Western blotting检测VASA在mRNA和蛋白水平的表达。结果：VASA mRNA在射出精子中有表达。QRT-PCR分析显示，正常精子症男性的VASA mRNA水平约为少精子症男性的5倍。免疫荧光和Western blotting分析表明，VASA蛋白定位于精子头部和尾部的胞质膜上，在少精子症患者中表达明显降低，与QRT-PCR结果相似。结论：VASA mRNA和蛋白在少精子症患者精子中的表达明显降低，提示VASA基因的低表达可能与某些类型的男性不育症的发病机制有关，VASA可作为诊断男性不育症的分子标志物。
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.