人类Izumo4和Tssk6在精子顶体反应前后的定位检测

Orientation of human Izumo4 and Tssk6 before and after sperm acrosome reaction

目的鉴定人Izumo4和Tssk6在精子顶体反应前后的位置。方法在实验室自制小鼠抗人Izumo4和Tssk6多克隆抗体,采用免疫组织化学法分别检测顶体反应前后Izumo4和Tssk6在人精子中所处的位置。结果顶体反应前,经抗HIS-Izumo4多克隆抗体处理的精子在头部赤道靠上部位染色稍浅,而赤道上膜染色稍深,经抗HIS-Tssk6多克隆抗体处理的精子在尾部和赤道靠上部位染色较深。顶体反应后,经抗HIS-Izumo4和抗HIS-Tssk6腹水处理过的精子顶体膜均明显破裂且对3-氨基-9-乙基咔唑染色剂不显色。结论 Tssk6位于精子头部的顶体内部及精子尾部,Izumo4位于顶体外膜上。

Objective To survey Izumo4 and Tssk6 positions before and after sperm acrosome reaction. Methods Anti-mouse polyelonal antibodies were made against Izumo4 and Tssk6 in our laboratory. Using im- munohistochemieal method, Izumo4 and Tssk6 positions in human sperms were detected before and after the acrosome reaction. Results Before the acrosome reaction, the sperms treated by anti-HIS-Izumo4 polyclonal antibody were stained light in the head above the equator, and a little deeper in the membrane on the equa- tor; those treated by anti-HIS-Tssk6 polyclonal antibody had deep stain in the tail and above the equator. After the aerosome reaction, the acrosome membrane was broken in the sperms treated with ascites containing anti-HIS-Izumo4 and anti-HIS-Tssk6 antibodies and eould not be stained by 3-amino-9-ethylcarbazole. Conclusions Tssk6 should be located in acrosome of sperm head and in sperm tail, while Izumo4 is located on the outer membrane of sperm acrosome.