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

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 secrete their content during cortical reaction. Here, we characterize cortic...

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Autores principales: Cappa, Andrea I., de Paola, Matilde, Wetten, Paula, De Blas, Gerardo A., Michaut, Marcela A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6310882/
https://www.ncbi.nlm.nih.gov/pubmed/30341105
http://dx.doi.org/10.1242/bio.031872
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author Cappa, Andrea I.
de Paola, Matilde
Wetten, Paula
De Blas, Gerardo A.
Michaut, Marcela A.
author_facet Cappa, Andrea I.
de Paola, Matilde
Wetten, Paula
De Blas, Gerardo A.
Michaut, Marcela A.
author_sort Cappa, Andrea I.
collection PubMed
description 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 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 (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 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.
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spelling pubmed-63108822018-12-31 Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content Cappa, Andrea I. de Paola, Matilde Wetten, Paula De Blas, Gerardo A. Michaut, Marcela A. Biol Open Research Article 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 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 (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 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. The Company of Biologists Ltd 2018-12-15 /pmc/articles/PMC6310882/ /pubmed/30341105 http://dx.doi.org/10.1242/bio.031872 Text en © 2018. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/4.0This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Research Article
Cappa, Andrea I.
de Paola, Matilde
Wetten, Paula
De Blas, Gerardo A.
Michaut, Marcela A.
Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
title Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
title_full Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
title_fullStr Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
title_full_unstemmed Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
title_short Live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
title_sort live imaging of cortical granule exocytosis reveals that in vitro matured mouse oocytes are not fully competent to secrete their content
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6310882/
https://www.ncbi.nlm.nih.gov/pubmed/30341105
http://dx.doi.org/10.1242/bio.031872
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