大鼠储精囊分泌蛋白Ⅱ在精子表面的荧光定位研究

FLUORESCENCE LOCALIZATION OF SEMINAL VESICLE SECRETION PROTEIN Ⅱ ON RAT SPERM SURFACE

本文报道应用免疫荧光定位技术首次证明大鼠储精囊分泌蛋白Ⅱ(SVPⅡ)和附睾精子表面结合。纯化的SVPⅡ和附睾精子一起培育后,经兔抗SVPⅡ抗血清和FITC标记的羊抗兔抗体共同保温,在荧光显微镜下观察,发现整个附睾精子表面被黄色的荧光所覆盖,且以精子头部为著;而同时以兔免疫前血清取代兔抗SVPⅡ抗血清作对照,精子表面未见任何荧光结合区。实验结果充分表明:SVPⅡ能和附睾精子表面呈特异结合,可能为大鼠精子包被抗原的成分。另外经免疫吸收试验和免疫交叉反应提示:SVPⅡ为组织特异抗原,与已纯化的其它SVP组分无交叉反应。

The binding of seminal vesicle secretion protein Ⅱ to rat epididymal sperm surfacewas studied by indirect immunofluorescent antibody technique. Rat epididymal sperms wereincubated with purified SVPⅡ then treated with rabbit anti-SVPⅡ serum and goat anti-rabbit IgG conjugated to fluorescein isothiocyanate, The sperm surface was completelycoated with fluorescence, and the fluorescence at the sperm head was brightest. In the controlgroup, the rabbit anti-SVPⅡ serum was replaced by the preimmune rabbit serum, nofluorescence was seen on sperm surface. It was brought to a conclusion that SVPⅡ hassome ability to bind the epididymal sperm surface and may be one of the components ofsperm coating antigen in rats. The immunoabsorption and immunocrossreaction tests furtherdemonstrated that SVPⅡ was tissue-specific antigen and there was no crossreaction withother purified SVP.