Cappa Andrea I, de Paola Matilde, Wetten Paula, De Blas Gerardo A, Michaut Marcela A
Laboratorio de Biología Reproductiva y Molecular, Instituto de Histología y Embriología, Universidad Nacional de Cuyo-CONICET, Av. Libertador 80, 5500, Mendoza, Argentina.
Universidad Nacional de Cuyo, Facultad de Ciencias Médicas, Área de Farmacología, Av. Libertador 80, 5500, Mendoza, Argentina.
Biol Open. 2018 Dec 3;7(12):bio031872. doi: 10.1242/bio.031872.
Oocyte maturation does not entirely support all the nuclear and cytoplasmic changes that occur physiologically, and it is poorly understood whether maturation affects the competence of cortical granules to secrete 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 ( matured, IVO) and 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 in regards to 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.This article has an associated First Person interview with the first author of the paper.
卵母细胞成熟并不完全支持生理上发生的所有核和细胞质变化,而且对于成熟是否会影响皮质颗粒在皮质反应过程中分泌其内容物的能力,人们了解甚少。在此,我们使用荧光凝集素FITC-LCA对氯化锶激活的活小鼠卵母细胞中的皮质颗粒胞吐作用(CGE)进行了表征。我们比较了排卵(成熟,IVO)和成熟(IVM)小鼠卵母细胞之间CGE的动力学。结果表明:(1)IVM卵母细胞对氯化锶的反应严重降低;(2)通过对通透细胞中残余皮质颗粒的定量以及一种用于定量非通透细胞中渗出物的新方法,证实了这种低反应;(3)IVO卵母细胞中CGE的动力学快速且同步;(4)IVM卵母细胞中CGE的动力学延迟且不同步;(5)IVM卵母细胞中的皮质颗粒在皮质颗粒自由域方面表现出不规则的界限。我们建议将活卵母细胞中CGE的分析作为一种生物学测试,以评估IVM小鼠卵母细胞的能力。本文配有对该论文第一作者的第一人称访谈。