Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secret their content

Abstract

Oocyte in vitro maturation does not entirely support all the nuclear and cytoplasmic changes that occur physiologically and it is poorly understood whether in vitro maturation affects the competence of cortical granules to secret their content during cortical reaction. Here, we characterize cortical granule exocytosis (CGE) in live mouse oocytes activated by strontium chloride using the fluorescent lectin FITC-LCA. We compared the kinetic of CGE between ovulated (in vivo matured, IVO) and in vitro matured (IVM) mouse oocytes. Results show that: 1) IVM oocytes have a severely reduced response to strontium chloride; 2) the low response was confirmed by quantification of remnant cortical granules in permeabilized cells and by a novel method to quantify the exudate in non permeabilized cells; 3) the kinetic of CGE in IVO oocytes was rapid and synchronous; 4) the kinetic of CGE in IVM oocytes was delayed and asynchronous; 5) cortical granules in IVM oocytes show an irregular limit with the cortical granule free domain. We propose the analysis of CGE in live oocytes as a biological test to evaluate the competence of IVM mouse oocytes.