Haneji T, Koide S S
Center for Biomedical Research, Population Council, New York, New York 10021.
Biol Reprod. 1987 Oct;37(3):675-83. doi: 10.1095/biolreprod37.3.675.
The serum obtained from an infertile woman induced a specific head-to-head agglutination of human and rat sperm. The immunoglobulin G (IgG) fraction of the serum was obtained and found to interact with the proteins of rat sperm in testis and epididymis. Using an indirect immunofluorescent method with rat sperm from vas deferens, we determined that the antibody recognized the protein on the convex and concave regions of the acrosome and over the entire tail. However, with testicular spermatozoa, the antibody recognized only the distal end of the tails. In paraffin sections of adult rat testis, sperm tails located at the luminal region of the seminiferous tubules stained intensely. Weak but significant staining also occurred on late spermatids. In the epididymal sections, staining was restricted to spermatozoa in the lumen. On the other hand, sections of testes from 25-day-old rats containing spermatogonia and early spermatocytes had a completely negative reaction. Testicular somatic cells, including Sertoli cells, peritubular myoid cells and interstitial cells, did not stain. To identify the testicular protein interacting with the antibody, adult rat testis proteins were prepared and analyzed by a sodium dodecyl sulfate-polyacrylamide gel electrophoretic (SDS-PAGE) immunoblot technique. The antibody interacted with a protein with an estimated molecular weight of 82,000 in the testicular homogenate and particulate fraction, whereas the reaction was considerably weaker with the testicular cytosol fraction.
从一名不孕妇女获得的血清可诱导人精子和大鼠精子发生特异性的头对头凝集。获取该血清的免疫球蛋白G（IgG）组分，发现其与睾丸和附睾中大鼠精子的蛋白质相互作用。使用来自输精管的大鼠精子进行间接免疫荧光法，我们确定该抗体识别顶体凸面和凹面区域以及整个尾部的蛋白质。然而，对于睾丸精子，该抗体仅识别尾部的远端。在成年大鼠睾丸的石蜡切片中，位于生精小管管腔区域的精子尾部染色强烈。晚期精子细胞也出现微弱但显著的染色。在附睾切片中，染色仅限于管腔中的精子。另一方面，来自25日龄大鼠的睾丸切片含有精原细胞和早期精母细胞，反应完全为阴性。睾丸体细胞，包括支持细胞、睾丸肌样细胞和间质细胞，均未染色。为了鉴定与该抗体相互作用的确睾丸蛋白，制备成年大鼠睾丸蛋白，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）免疫印迹技术进行分析。该抗体与睾丸匀浆和颗粒组分中一种估计分子量为82,000的蛋白质相互作用，而与睾丸胞质溶胶组分的反应则弱得多。
