Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs

Autores
de Paola, Maria Matilde; Bello, Oscar Daniel; Michaut, Marcela Alejandra
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Cortical granule exocytosis (CGE), also known as cortical reaction, is a calciumregulated secretion that represents a membrane fusion process during meiotic cell division of eggs. The molecular mechanism of membrane fusion during CGE is still poorly understood and is thought to be mediated by the SNARE pathway;nevertheless, it is unkown if SNAP (acronym for soluble NSF attachment protein) and NSF (acronym for N-ethilmaleimide sensitive factor), two key proteins in the SNARE pathway, mediate CGE in any egg model. In this paper, we documented the gene expression of α-SNAP, γ-SNAP and NSF in mouse oocytes. Western blot analysis showed that the expression of these proteins maintains a similar level during oocyte maturation and egg activation. Their localization was mainly observed at the cortical region of eggs, which is enriched in cortical granules. To evaluate the function of these proteins in CGE we set up a functional assay based on the quantification of cortical granules. Endogenous α-SNAP and NSF proteins were perturbed by microinjection of recombinant proteins or antibodies prior to CGE activation. The microinjection of wild type α-SNAP and the negative mutant of α-SNAP L294A in eggs inhibited CGE stimulated by strontium. NEM, an irreversibly inhibitor of NSF, and the microinjection ofthe negative mutant NSF D1EQ inhibited cortical reaction. The microinjection of anti-α-SNAP and anti-NSF antibodies was able to abolish CGE in activated eggs. The microinjection of anti-γ SNAP antibody had no effect on CGE. Our findings indicate, for the first time in any oocyte model, that α-SNAP, γ-SNAP, and NSF are expressed in mouse oocytes. We demonstrate that α-SNAP and NSF have an active role in CGE and propose a working model.
Fil: de Paola, Maria Matilde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo; Argentina
Fil: Bello, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. University of Yale; Estados Unidos
Fil: Michaut, Marcela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Materia
ALPHA-SNAP
NSF
GAMMA-SNAP
EXOCYTOSIS
CORTICAL REACTION
MOUSE OOCYTES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/41053
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/41053
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggsde Paola, Maria MatildeBello, Oscar DanielMichaut, Marcela AlejandraALPHA-SNAPNSFGAMMA-SNAPEXOCYTOSISCORTICAL REACTIONMOUSE OOCYTEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Cortical granule exocytosis (CGE), also known as cortical reaction, is a calciumregulated secretion that represents a membrane fusion process during meiotic cell division of eggs. The molecular mechanism of membrane fusion during CGE is still poorly understood and is thought to be mediated by the SNARE pathway;nevertheless, it is unkown if SNAP (acronym for soluble NSF attachment protein) and NSF (acronym for N-ethilmaleimide sensitive factor), two key proteins in the SNARE pathway, mediate CGE in any egg model. In this paper, we documented the gene expression of α-SNAP, γ-SNAP and NSF in mouse oocytes. Western blot analysis showed that the expression of these proteins maintains a similar level during oocyte maturation and egg activation. Their localization was mainly observed at the cortical region of eggs, which is enriched in cortical granules. To evaluate the function of these proteins in CGE we set up a functional assay based on the quantification of cortical granules. Endogenous α-SNAP and NSF proteins were perturbed by microinjection of recombinant proteins or antibodies prior to CGE activation. The microinjection of wild type α-SNAP and the negative mutant of α-SNAP L294A in eggs inhibited CGE stimulated by strontium. NEM, an irreversibly inhibitor of NSF, and the microinjection ofthe negative mutant NSF D1EQ inhibited cortical reaction. The microinjection of anti-α-SNAP and anti-NSF antibodies was able to abolish CGE in activated eggs. The microinjection of anti-γ SNAP antibody had no effect on CGE. Our findings indicate, for the first time in any oocyte model, that α-SNAP, γ-SNAP, and NSF are expressed in mouse oocytes. We demonstrate that α-SNAP and NSF have an active role in CGE and propose a working model.Fil: de Paola, Maria Matilde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo; ArgentinaFil: Bello, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. University of Yale; Estados UnidosFil: Michaut, Marcela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaPublic Library of Science2015-08-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/41053de Paola, Maria Matilde; Bello, Oscar Daniel; Michaut, Marcela Alejandra; Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs; Public Library of Science; Plos One; 10; 8; 12-8-2015; 1-191932-6203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0135679info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0135679info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:01:18Zoai:ri.conicet.gov.ar:11336/41053instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:01:18.953CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
title Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
spellingShingle Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
de Paola, Maria Matilde
ALPHA-SNAP
NSF
GAMMA-SNAP
EXOCYTOSIS
CORTICAL REACTION
MOUSE OOCYTES
title_short Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
title_full Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
title_fullStr Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
title_full_unstemmed Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
title_sort Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs
dc.creator.none.fl_str_mv de Paola, Maria Matilde
Bello, Oscar Daniel
Michaut, Marcela Alejandra
author de Paola, Maria Matilde
author_facet de Paola, Maria Matilde
Bello, Oscar Daniel
Michaut, Marcela Alejandra
author_role author
author2 Bello, Oscar Daniel
Michaut, Marcela Alejandra
author2_role author
author
dc.subject.none.fl_str_mv ALPHA-SNAP
NSF
GAMMA-SNAP
EXOCYTOSIS
CORTICAL REACTION
MOUSE OOCYTES
topic ALPHA-SNAP
NSF
GAMMA-SNAP
EXOCYTOSIS
CORTICAL REACTION
MOUSE OOCYTES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Cortical granule exocytosis (CGE), also known as cortical reaction, is a calciumregulated secretion that represents a membrane fusion process during meiotic cell division of eggs. The molecular mechanism of membrane fusion during CGE is still poorly understood and is thought to be mediated by the SNARE pathway;nevertheless, it is unkown if SNAP (acronym for soluble NSF attachment protein) and NSF (acronym for N-ethilmaleimide sensitive factor), two key proteins in the SNARE pathway, mediate CGE in any egg model. In this paper, we documented the gene expression of α-SNAP, γ-SNAP and NSF in mouse oocytes. Western blot analysis showed that the expression of these proteins maintains a similar level during oocyte maturation and egg activation. Their localization was mainly observed at the cortical region of eggs, which is enriched in cortical granules. To evaluate the function of these proteins in CGE we set up a functional assay based on the quantification of cortical granules. Endogenous α-SNAP and NSF proteins were perturbed by microinjection of recombinant proteins or antibodies prior to CGE activation. The microinjection of wild type α-SNAP and the negative mutant of α-SNAP L294A in eggs inhibited CGE stimulated by strontium. NEM, an irreversibly inhibitor of NSF, and the microinjection ofthe negative mutant NSF D1EQ inhibited cortical reaction. The microinjection of anti-α-SNAP and anti-NSF antibodies was able to abolish CGE in activated eggs. The microinjection of anti-γ SNAP antibody had no effect on CGE. Our findings indicate, for the first time in any oocyte model, that α-SNAP, γ-SNAP, and NSF are expressed in mouse oocytes. We demonstrate that α-SNAP and NSF have an active role in CGE and propose a working model.
Fil: de Paola, Maria Matilde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo; Argentina
Fil: Bello, Oscar Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. University of Yale; Estados Unidos
Fil: Michaut, Marcela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
description Cortical granule exocytosis (CGE), also known as cortical reaction, is a calciumregulated secretion that represents a membrane fusion process during meiotic cell division of eggs. The molecular mechanism of membrane fusion during CGE is still poorly understood and is thought to be mediated by the SNARE pathway;nevertheless, it is unkown if SNAP (acronym for soluble NSF attachment protein) and NSF (acronym for N-ethilmaleimide sensitive factor), two key proteins in the SNARE pathway, mediate CGE in any egg model. In this paper, we documented the gene expression of α-SNAP, γ-SNAP and NSF in mouse oocytes. Western blot analysis showed that the expression of these proteins maintains a similar level during oocyte maturation and egg activation. Their localization was mainly observed at the cortical region of eggs, which is enriched in cortical granules. To evaluate the function of these proteins in CGE we set up a functional assay based on the quantification of cortical granules. Endogenous α-SNAP and NSF proteins were perturbed by microinjection of recombinant proteins or antibodies prior to CGE activation. The microinjection of wild type α-SNAP and the negative mutant of α-SNAP L294A in eggs inhibited CGE stimulated by strontium. NEM, an irreversibly inhibitor of NSF, and the microinjection ofthe negative mutant NSF D1EQ inhibited cortical reaction. The microinjection of anti-α-SNAP and anti-NSF antibodies was able to abolish CGE in activated eggs. The microinjection of anti-γ SNAP antibody had no effect on CGE. Our findings indicate, for the first time in any oocyte model, that α-SNAP, γ-SNAP, and NSF are expressed in mouse oocytes. We demonstrate that α-SNAP and NSF have an active role in CGE and propose a working model.
publishDate 2015
dc.date.none.fl_str_mv 2015-08-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/41053
de Paola, Maria Matilde; Bello, Oscar Daniel; Michaut, Marcela Alejandra; Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs; Public Library of Science; Plos One; 10; 8; 12-8-2015; 1-19
1932-6203
CONICET Digital
CONICET
url http://hdl.handle.net/11336/41053
identifier_str_mv de Paola, Maria Matilde; Bello, Oscar Daniel; Michaut, Marcela Alejandra; Cortical granule exocytosis is mediated by alpha-SNAP and N-Ethilmaleimide sensitive factor in mouse eggs; Public Library of Science; Plos One; 10; 8; 12-8-2015; 1-19
1932-6203
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0135679
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0135679
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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