Haneji T, Koide S S
Center for Biomedical Research, The Population Council, New York, New York 10021.
Biol Reprod. 1987 Sep;37(2):467-77. doi: 10.1095/biolreprod37.2.467.
A monoclonal antibody (MAb) raised against human sperm protein, designated YWK-II, was used to determine the distribution of antigens in rat spermatozoa and rat testicular germ cells. By an indirect immunofluorescent method, the antibody localized over the rat spermatozoal head, except for the postacrosomal region. In paraffin sections of adult and immature rat testis, germ cells, at every developmental stage, and Sertoli cells stained, while interstitial cells and peritubular myoid cells remained unstained. When cocultures of Sertoli and germ cells were tested, only the germ cells stained intensely. Sertoli cells and peritubular myoid cells in cultures did not stain. In the epididymal sections, strong staining occurred with spermatozoa in the lumen and epididymal epithelial cells, with moderate staining in the myoid layers of epididymis. To determine the sperm antigen interacting with the YWK-II antibody, rat spermatozoa proteins were prepared and analyzed by an immunoblot technique. The monoclonal antibody interacted with a single protein, with an estimated molecular weight of 115,000, present in the cauda epididymal spermatozoa. Among the proteins of the caput epididymal spermatozoa, however, the antibody interacted with a major and a minor band with molecular weights of 115,000 and 88,000, respectively. On the other hand, with proteins prepared from the membrane fraction of adult and immature rat testis, the antibody reacted with two bands with estimated molecular weights of 88,000 and 115,000. In the lysate prepared from germ cells dissociated from Sertoli-germ cell cocultures, the antibody recognized only the 88,000 protein. The present results show that the YWK-II MAb interacts with two proteins with different molecular weights. The amount of the interacting proteins in spermatozoa varied with their location within the epididymis.
一种针对人精子蛋白产生的单克隆抗体（MAb），命名为YWK-II，用于确定抗原在大鼠精子和大鼠睾丸生殖细胞中的分布。通过间接免疫荧光法，该抗体定位于大鼠精子头部，但顶体后区域除外。在成年和未成熟大鼠睾丸的石蜡切片中，各个发育阶段的生殖细胞和支持细胞均被染色，而间质细胞和睾丸肌样细胞未被染色。当检测支持细胞与生殖细胞的共培养物时，只有生殖细胞被强烈染色。培养物中的支持细胞和睾丸肌样细胞未被染色。在附睾切片中，管腔内的精子和附睾上皮细胞出现强染色，附睾肌样层呈中度染色。为了确定与YWK-II抗体相互作用的精子抗原，制备了大鼠精子蛋白并通过免疫印迹技术进行分析。该单克隆抗体与一种单一蛋白质相互作用，估计分子量为115,000，存在于附睾尾精子中。然而，在附睾头精子的蛋白质中，该抗体与分子量分别为115,000和88,000的一条主要带和一条次要带相互作用。另一方面，对于从成年和未成熟大鼠睾丸膜部分制备的蛋白质，该抗体与估计分子量为88,000和115,000的两条带发生反应。在从支持细胞 - 生殖细胞共培养物中解离的生殖细胞制备的裂解物中，该抗体仅识别88,000的蛋白质。目前的结果表明，YWK-II单克隆抗体与两种分子量不同的蛋白质相互作用。精子中相互作用蛋白质的量随其在附睾内的位置而变化。
