Cherr G N, Drobnis E Z, Katz D F
Bodega Marine Laboratory, Bodega Bay, California 94923.
J Exp Zool. 1988 Apr;246(1):81-93. doi: 10.1002/jez.1402460111.
Electrical activation of the hamster egg was used to study cortical granule constituents before and after exocytosis. The activated hamster eggs underwent cortical granule decondensation just prior to and at the time of exocytosis. Some of the cortical granules of aged, unactivated eggs underwent similar changes. FITC- and gold-conjugated Lens culinaris agglutinin (LCA) bound intensely to the surfaces of activated but not unactivated eggs. This labelling was associated with the microvilli. Permeabilized eggs exhibited discrete cortical labelling before activation, with a subsequent decrease following the cortical reaction. Gold-conjugated LCA specifically bound to cortical granules when incubated with thin sections. FITC-soybean trypsin inhibitor (SBTI) bound in discrete foci in the cortex of unactivated eggs. Following activation, cortical labelling by SBTI decreased. Aprotinin and benzamidine hydrochloride inhibited FITC-SBTI from binding to the egg cortex. Gold-avidin localization of biotin-SBTI in the electron microscope demonstrated that condensed cortical granules did not bind SBTI but decondensed or exocytosing granules did. This suggests that a cortical granule protease is exposed just prior to exocytosis. Activated eggs exhibited dramatic decreases in the number of hamster sperm penetrating the cytoplasm, suggesting that a plasma membrane block to polyspermy is temporally related to cortical granule exocytosis.
通过电激活仓鼠卵来研究皮质颗粒成分在胞吐作用前后的变化。激活后的仓鼠卵在胞吐作用之前及之时会发生皮质颗粒解聚。一些老龄未激活卵的部分皮质颗粒也会发生类似变化。异硫氰酸荧光素(FITC)和金标记的菜豆凝集素(LCA)强烈结合于激活卵而非未激活卵的表面。这种标记与微绒毛有关。通透处理的卵在激活前呈现离散的皮质标记,皮质反应后标记减少。金标记的LCA与薄片孵育时可特异性结合皮质颗粒。FITC - 大豆胰蛋白酶抑制剂(SBTI)在未激活卵的皮质中以离散位点结合。激活后,SBTI的皮质标记减少。抑肽酶和盐酸苯甲脒可抑制FITC - SBTI与卵皮质结合。电子显微镜下生物素 - SBTI的金抗生物素蛋白定位显示,浓缩的皮质颗粒不结合SBTI,但解聚或正在胞吐的颗粒会结合。这表明皮质颗粒蛋白酶在胞吐作用即将发生前暴露。激活的卵穿透细胞质的仓鼠精子数量显著减少,这表明质膜对多精受精的阻断在时间上与皮质颗粒胞吐作用相关。