Egawa Miho, Yoshioka Shinya, Higuchi Toshihiro, Sato Yukiyasu, Tatsumi Keiji, Fujiwara Hiroshi, Fujii Shingo
Department of Gynecology and Obstetrics, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8507, Japan.
J Clin Endocrinol Metab. 2003 Sep;88(9):4384-92. doi: 10.1210/jc.2002-021910.
Ephrins and their Eph receptors are both membrane-bound proteins that function in various cell-cell recognition processes, such as morphogenesis and angiogenesis. In this study we examined the expression of B class ephrins-Ephs in the human ovary during corpus luteum formation, a process of tissue remodeling accompanied by angiogenesis. RT-PCR analysis detected mRNAs for Eph B1, B2, and B4 and ephrin B1 and B2, but not Eph B3 and B6 or ephrin B3, in human corpora lutea of the early luteal phase. By immunohistochemistry, ephrin B1 was moderately expressed on theca interna cells, but was expressed at a low level on granulosa cells in the preovulatory follicles. After ovulation, a rapid increase in ephrin B1 expression was observed on luteinizing granulosa cells, whereas its expression on luteinizing theca interna cells decreased. The mRNA expression of ephrin B1 in luteinizing granulosa cells was confirmed by Northern blotting. Flow cytometry showed that ephrin B1 was expressed on the surface of isolated luteinizing granulosa cells. Moreover, these cells had the ability to bind to recombinant Eph B2-Fc fusion protein. These findings suggest that ephrin B1-expressing granulosa cells can directly interact with Eph-bearing cells during corpus luteum formation in vivo, suggesting that Eph-ephrin system is involved in this process.
Ephrin 及其 Eph 受体均为膜结合蛋白,在多种细胞间识别过程中发挥作用,如形态发生和血管生成。在本研究中,我们检测了 B 类 Ephrin - Eph 在人卵巢黄体形成过程中的表达,黄体形成是一个伴随着血管生成的组织重塑过程。RT - PCR 分析在黄体早期的人黄体中检测到了 Eph B1、B2 和 B4 以及 ephrin B1 和 B2 的 mRNA,但未检测到 Eph B3 和 B6 或 ephrin B3 的 mRNA。通过免疫组织化学方法,发现 ephrin B1 在卵泡膜内层细胞中中度表达,但在排卵前卵泡的颗粒细胞中表达水平较低。排卵后,在黄素化颗粒细胞上观察到 ephrin B1 表达迅速增加,而在黄素化卵泡膜内层细胞上的表达则下降。Northern 印迹法证实了黄素化颗粒细胞中 ephrin B1 的 mRNA 表达。流式细胞术显示 ephrin B1 在分离的黄素化颗粒细胞表面表达。此外,这些细胞具有与重组 Eph B2 - Fc 融合蛋白结合的能力。这些发现表明,在体内黄体形成过程中,表达 ephrin B1 的颗粒细胞可直接与携带 Eph 的细胞相互作用,提示 Eph - ephrin 系统参与了这一过程。