Yanaihara A, Yanaihara T, Toma Y, Shimizu Y, Saito H, Okai T, Higashiyama T, Osawa Y
Department of Obstetrics and Gynecology, Showa University School of Medicine, 1-5-8, Hatanodai, Shinagawa-ku, 142-8666, Tokyo, Japan.
Steroids. 2001 Feb;66(2):87-91. doi: 10.1016/s0039-128x(00)00204-x.
Localization of steroid sulfatase, a membrane-bound microsomal enzyme, in human fallopian tubes was immunohistochemically investigated, and expression of RNA was confirmed by competitive RT-PCR. Human fallopian tubes were obtained from 10 patients in follicular and early luteal phases during gynecological laparotomy. An anti-human rabbit polyclonal antibody was prepared against sulfatase protein purified from human placenta. Total RNA was isolated from epithelium of fallopian tubes. A heterologous RNA competitor was designed, and competitive RT-PCR was carried out. Steroid sulfatase was localized to the cytoplasm of epithelial cells. With respect to the positive staining of cells, the number of positive secretory cells was higher than that of ciliated cells. A significantly higher number of positive cells was found in tissue obtained from the early luteal phase than that found in tissue from the follicular phase. An abundant expression of sulfatase mRNA in early luteal phase was also observed. This study demonstrates, for the first time, that steroid sulfatase is localized to human epithelial cells and that steroid sulfatase staining and mRNA expression changes with the menstrual cycle. These results suggest that sulfatase in the fallopian tube may be involved in controlling the local steroid environment, which appears to regulate aspects of the physiological reproductive function of the fallopian tube.
采用免疫组织化学方法研究了膜结合微粒体酶类固醇硫酸酯酶在人输卵管中的定位,并通过竞争性逆转录聚合酶链反应(RT-PCR)证实了其RNA的表达。从10例接受妇科剖腹手术的患者卵泡期和黄体早期的输卵管中获取样本。针对从人胎盘中纯化的硫酸酯酶蛋白制备了抗人兔多克隆抗体。从输卵管上皮中分离总RNA。设计了一种异源RNA竞争物,并进行了竞争性RT-PCR。类固醇硫酸酯酶定位于上皮细胞的细胞质中。就细胞的阳性染色而言,阳性分泌细胞的数量高于纤毛细胞。在黄体早期获得的组织中发现的阳性细胞数量明显高于卵泡期组织中的阳性细胞数量。在黄体早期还观察到硫酸酯酶mRNA的丰富表达。本研究首次证明类固醇硫酸酯酶定位于人上皮细胞,并且类固醇硫酸酯酶染色和mRNA表达随月经周期而变化。这些结果表明,输卵管中的硫酸酯酶可能参与控制局部类固醇环境,这似乎调节了输卵管生理生殖功能的各个方面。