论文部分内容阅读
用高度纯化的小鼠睾丸乳酸脱氢酶C_4(LDH-C_4)免疫家兔,获得兔抗小鼠LDH-C_4抗血清。以1:20稀释的兔抗小鼠LDH-C_4抗体为第一抗体,1:10稀释的羊抗兔IgG荧光抗体为第二抗体,对小鼠精子和人精子的LDH-C_4用免疫荧光法进行定位。人精子取自有生育力男性的新鲜精液;小鼠精子取自成年小鼠的附睾,新鲜小鼠睾丸制片为4μm冷冻切片。免疫荧光定位表明,小鼠和人精子的表面有较强的荧光结合,提示精子表面有LDH-C_4分子存在。人精子的LDH-C_4主要定位于颈部和中段;而在小鼠精子,荧光抗体主要结合于尾部主段。酶学组织化学定位研究结果表明,在人和小鼠精子的颈部和中段,以及靠近小鼠曲细精管管腔的细胞中,有颜色较深的甲(月朁)沉淀。小鼠精子的免疫荧光定位结果与酶组织化学染色结果不一致,其原因有待探讨。
Rabbit anti-mouse LDH-C_4 antiserum was obtained by immunizing rabbits with highly purified mouse testis lactate dehydrogenase C_4 (LDH-C_4). The rabbit anti-mouse LDH-C_4 antibody diluted 1:20 was the first antibody, the goat anti-rabbit IgG fluorescent antibody 1:10 diluted was the second antibody, and the LDH-C_4 of mouse sperm and human sperm was immunofluorescence Positioning. Human sperm were taken from fresh sperm of fertile men; mouse sperm were taken from the epididymis of adult mice; fresh mouse testes were frozen at 4 μm. Immunofluorescence staining showed that there was a strong fluorescence on the surface of mouse and human sperm, indicating that LDH-C_4 was present on the surface of sperm. Human sperm LDH-C_4 mainly located in the neck and middle; while in mouse sperm, fluorescent antibody mainly in the tail section. Enzymatic histochemical localization studies showed dark-colored formazan deposits in the neck and middle of human and mouse spermatozoa and in cells near the seminiferous tubules of mice. Mouse sperm immunofluorescence localization results and enzyme histochemical staining results do not agree, the reason to be explored.