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Compensatory endocytosis occurs after cortical granule exocytosis in mouse eggs
Authors:Matías D. Gómez-Elías  Rafael A. Fissore  Patricia S. Cuasnicú  Débora J. Cohen
Affiliation:1. Laboratorio de Mecanismos Moleculares de la Fertilización, Instituto de Biología y Medicina Experimental, Consejo Nacional de Investigaciones Científico y Técnicas (IBYME-CONICET), Buenos Aires, Argentina;2. Department of Veterinary and Animal Sciences, University of Massachusetts Amherst, 661 North Pleasant Street, Amherst, Massachusetts
Abstract:Compensatory endocytosis (CE) is one of the primary mechanisms through which cells maintain their surface area after exocytosis. Considering that in eggs massive exocytosis of cortical granules (CG) takes place after fertilization, the aim of this study was to evaluate the occurrence of CE following cortical exocytosis in mouse eggs. For this purpose, we developed a pulse-chase assay to detect CG membrane internalization. Results showed internalized labeling in SrCl2-activated and fertilized eggs when chasing at 37°C, but not at a nonpermissive temperature (4°C). The use of kinase and calcineurin inhibitors led us to conclude that this internal labeling corresponded to CE. Further experiments showed that CE in mouse eggs is dependent on actin dynamics and dynamin activity, and could be associated with a transient exposure of phosphatidylserine. Finally, CE was impaired in A23187 ionophore-activated eggs, highlighting once again the mechanistic differences between the activation methods. Altogether, these results demonstrate for the first time that egg activation triggers CE in mouse eggs after exocytosis of CG, probably as a plasma membrane homeostasis mechanism.
Keywords:compensatory endocytosis  cortical granules  egg activation  fertilization  phosphatidylserine
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