Bement W M, Benink H, Mandato C A, Swelstad B B
Department of Zoology, University of Wisconsin, Madison, Wisconsin 53706, USA.
J Exp Zool. 2000 Jun 1;286(7):767-75. doi: 10.1002/(sici)1097-010x(20000601)286:7<767::aid-jez11>3.0.co;2-l.
Rapid exocytosis is typically followed by rapid resorption of exocytosed membrane; however, whether membrane retrieval occurs via indirect endocytosis of numerous small vesicles or direct resealing of the original, larger exocytotic vesicles is controversial. Here we show that cortical granule (CG) exocytosis in Xenopus oocytes and eggs is followed by rapid formation of endosomes as large as the CGs. Large endosomes are translucent, and their formation has the same developmental and pharmacological profile as CG exocytosis. Time course analyses show that large endosomes are not derived from small endosomes. Large endosome formation is triggered by stimuli that do not trigger increases in intracellular-free calcium and is insensitive to perturbation of microtubules by treatment with nocodazole. Perturbation of the f-actin cytoskeleton with latrunculin, however, sharply reduces large endosome formation. We conclude that CG membrane is directly retrieved in Xenopus oocytes and eggs and suggest that this retrieval is not directly dependent on an increase in intracellular-free calcium, but is dependent on the actin cytoskeleton.
快速胞吐作用之后通常紧接着是胞吐膜的快速重吸收;然而,膜回收是通过众多小囊泡的间接内吞作用还是原始较大胞吐囊泡的直接重新封闭发生,这存在争议。在这里我们表明,非洲爪蟾卵母细胞和卵中的皮质颗粒(CG)胞吐作用之后会迅速形成与CG一样大的内体。大型内体是半透明的,它们的形成与CG胞吐作用具有相同的发育和药理学特征。时间进程分析表明,大型内体并非源自小型内体。大型内体的形成由不会引发细胞内游离钙增加的刺激触发,并且对用诺考达唑处理引起的微管扰动不敏感。然而,用拉冬库林扰乱f - 肌动蛋白细胞骨架会大幅减少大型内体的形成。我们得出结论,非洲爪蟾卵母细胞和卵中的CG膜是直接回收的,并表明这种回收不直接依赖于细胞内游离钙的增加,而是依赖于肌动蛋白细胞骨架。