Abe Hiroyuki, Kinoh Hiroaki, Oikawa Taneaki, Suzuki Norio
Research Institute for the Functional Peptides, 990, Minamisanbancho, Yamagata, Japan.
Noto Marine Laboratory, Kanazawa University, Ogi, Uchiura, 927-05, Ishikawa, Japan.
Rouxs Arch Dev Biol. 1992 May;201(3):179-189. doi: 10.1007/BF00188717.
The immunocytochemical localization of the egg-jelly macromolecule, a fucose sulphate glycoconjugate (FSG) that induces the acrosome reaction in spermatozoa, was investigated in ovaries of the sea urchin Hemicentrotus pulcherrimus by use of a polyclonal antibody. The polyclonal antibody reacted with the accessory cells and oocytes in the ovarian lumen. In the accessory cells, evidence of an intense immunohistochemical reaction was observed in many globules of variable density. Products of the specific immunohistochemical reaction were frequently observed in the surface region of oocytes, at a distance from the ovarian wall. At the ultrastructural level, the polyclonal antibody was found to react with the material present in the vacuole-like structures of the globules in the accessory cells. Many gold particles, demonstrating specific immunolabelling, were associated with well-developed microvilli on the vitellogenic oocytes. In the mature oocytes, intense labelling was observed in the jelly coat but not in the vitelline coat. By contrast, oogonia and early oocytes were barely labelled. Quantitative data indicated that the extent of immunolabellings in the surface region of oocytes was very high in the vitellogenic and mature oocytes. In all cases, neither the oocyte cytoplasm nor the subcellular organelles were labelled. These results suggest that FSG is produced by the accessory cells and is deposited initially on the surface of vitellogenic oocytes for the formation of jelly. These findings may provide a new insight into the role of the accessory cells in the reproductive process of the sea urchin.
利用多克隆抗体,在海胆光棘球海胆的卵巢中研究了卵胶大分子(一种能诱导精子顶体反应的岩藻糖硫酸糖缀合物(FSG))的免疫细胞化学定位。多克隆抗体与卵巢腔内的辅助细胞和卵母细胞发生反应。在辅助细胞中,在许多密度可变的小球中观察到强烈免疫组化反应的迹象。在离卵巢壁一定距离的卵母细胞表面区域经常观察到特异性免疫组化反应的产物。在超微结构水平上,发现多克隆抗体与辅助细胞中小球的液泡样结构中存在的物质发生反应。许多显示特异性免疫标记的金颗粒与卵黄生成期卵母细胞上发育良好的微绒毛相关。在成熟卵母细胞中,在卵胶膜中观察到强烈标记,但在卵黄膜中未观察到。相比之下,卵原细胞和早期卵母细胞几乎没有标记。定量数据表明,在卵黄生成期和成熟卵母细胞中,卵母细胞表面区域的免疫标记程度非常高。在所有情况下,卵母细胞细胞质和亚细胞器均未被标记。这些结果表明,FSG由辅助细胞产生,并最初沉积在卵黄生成期卵母细胞表面以形成卵胶。这些发现可能为辅助细胞在海胆生殖过程中的作用提供新的见解。