牛精子头后部质膜分区的研究

TWO REGIONS ON POSTERIOR HEAD OF BULL SPERMATOZOA

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摘要目的检测牛精子头后部存在的不同质膜区域。方法先用钙离子载体Ａ２３１８７诱导精子发生顶体反应，然后用水浴超声波和低速离心法分离得到纯牛精子头。直接用顶体反应后精子和低渗处理的顶体反应后精子头免疫Ｂａｌｂ／ｃ母鼠和新西兰公兔，制备抗牛精子抗体，并结合以前制备的抗牛精子ｓｐ１８单克隆抗体进行间接免疫组织化学分析。结果牛精子头后部质膜表面存在两个位置互补的荧光区域，即紧邻赤道板和紧邻尾段的荧光区域：前一区域表面稳定存在的主要是ｓｐ１８族膜蛋白，后者的膜蛋白在顶体反应和洗涤时易解离。结论依据膜蛋白成分的差异，牛精子头后部质膜可以划分为紧邻赤道板的ｓｐ１８质膜区域（ｓｐ１８－ｒｉｃｈｒｅｇｉｏｎ）和紧邻尾段的无ｓｐ１８质膜区域（ｓｐ１８－ｆｒｅｅｒｅｇｉｏｎ）。 Objective\ To study different domains on posterior head of bull spermatozoa.Methods Acrosome reacted bovine spermatozoa heads(A 23187 induced)were separated from their tails with ultrasonic treatment and centrifugation.Anti bovine spermatozoa IgG was produced by immunizing Balb/c mice or rabbit with acrosome reacted bovine spermatozoon or hypotonically treated acrosome reacted bovine spermatozoa heads.Results Indirect immunofluorescence microscopy showed that there were two complementary fluorescence regions on posterior head of the thawed bovine spermatozoa.One was adjacent to the equatorial segment and the other adjacent to the spermatozoon tail.They had different membrane proteins on two regions.The mAb to sp18 antigen was bound primarily to the region adjacent to the equatorial segment.Conclusion The posterior head of bovine spermatozoa divided into sp18 rich region adjacent to the equatorial segment and sp18 free region adjacent to the tail.There was no effect of ultrasonic treatment on membrance proteins on sp18 rich region and the membrane proteins on sp18 free region were easily to loss during acrosome reaction.