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目的探讨生殖细胞高度特异性基因VASA和信号分子BMP-4在精子发生中的作用和相关机制。方法正常和少精子症患者精液通过Percoll梯度分离获得精子和未成熟生精细胞,采用RT-PCR、real-time PCR、Western Blot和免疫荧光方法分析基因转录产物表达。结果少精子症精子VASA基因转录产物是正常精子的1/5；两种样本精子中均检测到荧光信号,正常组信号强,与Western blot结果相一致；两种样本精子RT-PCR均未检测到BMP-4及其Ⅰ型受体Alk-3表达,但在分离的未成熟生精细胞中均表达。人正常睾丸组织可检测到BMP- 4及其Alk-3表达。结论少精子症患者精子发生过程中VASA基因及其产物发生变化；BMP-4及其受体Alk- 3并不直接上调VASA表达；睾丸组织包括精子所存在的部分转录产物,而精子转录产物并不完全等同于睾丸组织；睾丸组织内表达BMP-4和Alk-3的可能主要是精原和精母细胞。
Objective To investigate the role and mechanism of VASA, a gene specific for germ cells, and BMP-4, a signaling molecule, in spermatogenesis. Methods Spermatozoa and immature spermatogenic cells were obtained from semen of normal and oligospermia patients by Percoll gradient. The expression of gene transcripts was analyzed by RT-PCR, real-time PCR, Western Blot and immunofluorescence. Results Sperm oligospermia sperm VASA gene transcripts were normal sperm 1/5; both samples were detected in the fluorescence signal of sperm, the signal of the normal group, consistent with the results of Western blot; two samples of sperm RT-PCR were not detected To BMP-4 and its type I receptor Alk-3, but expressed in isolated immature spermatogenic cells. Human normal testicular tissue can detect BMP-4 and its Alk-3 expression. Conclusion The expression of VASA gene and its products in spermatogenesis of spermatogenesis is changed. BMP-4 and its receptor Alk-3 do not directly up-regulate the expression of VASA. The testicular tissues include some transcription products present in spermatozoa, Not exactly equivalent to testicular tissue; testicular tissue expression of BMP-4 and Alk-3 may be mainly spermatogonia and spermatocytes.