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目的:探讨受激活调节正常T细胞表达和分泌因子(RANTES)在小鼠附睾与精子的结合情况。方法:应用免疫荧光双标法检测RANTES在小鼠附睾中的细胞定位;用RT-PCR方法确定小鼠附睾精子是否表达RANTES;用流式细胞技术观察RANTES与附睾各段精子的结合情况;用免疫荧光方法显示RANTES及其特异性受体CCR1在小鼠附睾精子上的定位。结果:RANTES在小鼠附睾组织中主要表达于狭窄细胞和顶细胞;不表达于小鼠附睾精子,但能与小鼠附睾精子结合;其与精子结合比例在小鼠附睾的头部最高,体部较少,尾部降至最低;并证实RANTES定位于精子的尾部中段,而受体CCR1定位于精子的头部。结论:RANTES可能在小鼠附睾头部参与了精子成熟过程的调节,RANTES特异性结合于精子尾部中段可能与精子运动相关;而RANTES与CCR1在小鼠附睾精子上的定位不一致,提示RANTES是通过CCR1以外的途径与精子结合。
Objective: To investigate the binding of activated and regulated normal T cells expression and secretion factor (RANTES) in mouse epididymis and sperm. METHODS: The cell localization of RANTES in mouse epididymis was detected by immunofluorescence double-labeled method. The expression of RANTES in mouse epididymal sperm was detected by RT-PCR. The binding of RANTES to each segment of sperm in epididymis was observed by flow cytometry. Immunofluorescence method showed the localization of RANTES and its specific receptor CCR1 in mouse epididymal sperm. RESULTS: RANTES was expressed mainly in the spermatocytes and the apical cells in the epididymis of mice; it was not expressed in the epididymal sperm of mice but could bind to the sperm of mouse epididymis; the ratio of sperm to spermatozoa was the highest in the epididymis of mice Less, tail to a minimum; and confirmed RANTES located in the sperm middle tail, and the receptor CCR1 located in the sperm head. Conclusion: RANTES may play an important role in the process of sperm maturation in the mouse epididymis head. RANTES specific binding to the sperm midpoint may be related to sperm motility. RANTES and CCR1 are located in the mouse epididymis inconsistent, suggesting that RANTES is through Pathways other than CCR1 bind to sperm.