Russinova A I, Valkova C A, Denkova R T
Institute of Experimental Morphology and Anthropology, Department of Cell Differentiation, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria.
Endocr Regul. 2001 Sep;35(3):127-37.
In the present study we employed a monoclonal antibody (Mab 3D8) obtained against a rat ovarian antigen and identified a 76 kDa protein in porcine ovarian follicles.
The localization of this antigen was studied by light and electron-microscopic immunocytochemistry and further characterized by polyacrylamide gel electrophoresis and immunoblotting on nitrocellulose membranes.
We found that the antigen recognized by Mab 3D8 is localized in granulosa cells (GCs) and oocytes. The expression of the 76 kDa protein apparently depends on the developmental stage. A particularly strong reaction was observed in cumulus cells and oocytes in early and late antral follicles. In granulosa cells the reaction product was localized in rough endoplasmic reticulum (RER), cis and trans faces of the Golgi stack, the outer nuclear envelope and in numerous transport vesicles budding from the endoplasmic reticulum. In the oocyte the reaction product was localized in structures related to specific endocytosis - small pits at the cell surface, two subsets of endosomes, endocytic carrier vesicles and the prelysosomal compartment (PLC).
The results obtained suggest that porcine oocytes possess the cellular structures, which allow them to bind and internalize this protein, which is most probably produced by granulosa cells. During oocyte development the intracellular site of accumulation of the 76 kDa protein varies, which implies that it is under developmental control. follicle
在本研究中，我们使用了一种针对大鼠卵巢抗原获得的单克隆抗体（Mab 3D8），并在猪卵巢卵泡中鉴定出一种76 kDa的蛋白质。
通过光镜和电镜免疫细胞化学研究该抗原的定位，并通过聚丙烯酰胺凝胶电泳和硝酸纤维素膜免疫印迹进一步表征。
我们发现Mab 3D8识别的抗原定位于颗粒细胞（GCs）和卵母细胞中。76 kDa蛋白质的表达显然取决于发育阶段。在早期和晚期窦状卵泡的卵丘细胞和卵母细胞中观察到特别强烈的反应。在颗粒细胞中，反应产物定位于粗面内质网（RER）、高尔基体堆栈的顺面和反面、外核膜以及从内质网出芽的许多运输小泡中。在卵母细胞中，反应产物定位于与特定内吞作用相关的结构——细胞表面的小窝、两种内体亚群、内吞载体小泡和前溶酶体区室（PLC）。
获得的结果表明，猪卵母细胞具有允许它们结合并内化这种最有可能由颗粒细胞产生的蛋白质的细胞结构。在卵母细胞发育过程中，76 kDa蛋白质的细胞内积累位点会发生变化，这意味着它受发育控制。卵泡
