Russinova A, Vassilev A, Davidoff M
Institute of Cell Biology and Morphology, Bulgarian Academy of Sciences, Sofia.
Biol Cell. 1993;79(3):259-64. doi: 10.1016/0248-4900(93)90145-5.
Hybridoma cell lines were obtained from mouse splenocytes sensitized to granulosa cells collected from rat ovaries after gonadotropin stimulation. A monoclonal antibody (5G5) was obtained which reacted with granulosa cells and showed a positive reaction with serum-free conditioned medium containing granulosa cell secreted proteins. Immunoblotting of the conditioned medium and light- and electron-microscopic immunocytochemistry of rat ovary show that mAb 5G5 is directed against a 59-kDa protein which is located on the plasma membrane of granulosa cells. Furthermore, the immunoreactivity of the granulosa cells depends both on the degree of follicle development and on the position of the granulosa cells within the follicles. Strong immunoreactivity was observed in the innermost granulosa cell layers, close to the oocyte and the antral cavity. The results obtained show that mAb 5G5 is a useful marker of a 59-kDa granulosa cell protein which might be of importance for the development of the follicle and the oocyte maturation.
杂交瘤细胞系源自对促性腺激素刺激后从大鼠卵巢收集的颗粒细胞致敏的小鼠脾细胞。获得了一种单克隆抗体（5G5），它与颗粒细胞发生反应，并与含有颗粒细胞分泌蛋白的无血清条件培养基呈阳性反应。对条件培养基进行免疫印迹以及对大鼠卵巢进行光镜和电镜免疫细胞化学分析表明，单克隆抗体5G5针对的是一种位于颗粒细胞质膜上的59 kDa蛋白质。此外，颗粒细胞的免疫反应性既取决于卵泡发育程度，也取决于颗粒细胞在卵泡内的位置。在最内层的颗粒细胞层，靠近卵母细胞和卵泡腔处观察到强免疫反应性。所获得的结果表明，单克隆抗体5G5是一种59 kDa颗粒细胞蛋白的有用标志物，该蛋白可能对卵泡发育和卵母细胞成熟具有重要意义。
