Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3

Autores
Maldera, Julieta Antonella; Weigel Muñoz, Mariana; Chirinos, M.; Busso, Dolores; Raffo, F. G. E.; Battistone, Maria Agustina; Blaquier, J. A.; Larrea, F.; Cuasnicu, Patricia Sara
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm-ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm?egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein?protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved notonly in sperm?egg fusion but also in the previous stage of sperm?ZP binding through its specific interaction with human ZP3.
Fil: Maldera, Julieta Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Heidelberg University. Center for Molecular Biology; Alemania
Fil: Weigel Muñoz, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Chirinos, M.. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran; México
Fil: Busso, Dolores. Pontificia Universidad Católica de Chile; Chile. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Raffo, F. G. E.. Centro Médico Fertilab; Argentina
Fil: Battistone, Maria Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Blaquier, J. A.. Centro Médico Fertilab; Argentina
Fil: Larrea, F.. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran; México
Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Materia
FERTILIZATION
HCRISP1
OOCYTE
SPERM
ZONA PELLUCIDA
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:11336/6291
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/6291
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3Maldera, Julieta AntonellaWeigel Muñoz, MarianaChirinos, M.Busso, DoloresRaffo, F. G. E.Battistone, Maria AgustinaBlaquier, J. A.Larrea, F.Cuasnicu, Patricia SaraFERTILIZATIONHCRISP1OOCYTESPERMZONA PELLUCIDAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm-ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm?egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein?protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved notonly in sperm?egg fusion but also in the previous stage of sperm?ZP binding through its specific interaction with human ZP3.Fil: Maldera, Julieta Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Heidelberg University. Center for Molecular Biology; AlemaniaFil: Weigel Muñoz, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Chirinos, M.. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran; MéxicoFil: Busso, Dolores. Pontificia Universidad Católica de Chile; Chile. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Raffo, F. G. E.. Centro Médico Fertilab; ArgentinaFil: Battistone, Maria Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Blaquier, J. A.. Centro Médico Fertilab; ArgentinaFil: Larrea, F.. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran; MéxicoFil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaOxford University Press2014-04-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/6291Maldera, Julieta Antonella; Weigel Muñoz, Mariana; Chirinos, M.; Busso, Dolores; Raffo, F. G. E.; et al.; Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3; Oxford University Press; Molecular Human Reproduction; 20; 4; 1-4-2014; 341-3491360-99471460-2407enginfo:eu-repo/semantics/altIdentifier/doi/info:eu-repo/semantics/altIdentifier/url/http://molehr.oxfordjournals.org/content/20/4/341.longinfo:eu-repo/semantics/altIdentifier/doi/10.1093/molehr/gat092info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:37:46Zoai:ri.conicet.gov.ar:11336/6291instacron: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:34982025-09-29 10:37:47.142CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
title Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
spellingShingle Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
Maldera, Julieta Antonella
FERTILIZATION
HCRISP1
OOCYTE
SPERM
ZONA PELLUCIDA
title_short Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
title_full Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
title_fullStr Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
title_full_unstemmed Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
title_sort Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3
dc.creator.none.fl_str_mv Maldera, Julieta Antonella
Weigel Muñoz, Mariana
Chirinos, M.
Busso, Dolores
Raffo, F. G. E.
Battistone, Maria Agustina
Blaquier, J. A.
Larrea, F.
Cuasnicu, Patricia Sara
author Maldera, Julieta Antonella
author_facet Maldera, Julieta Antonella
Weigel Muñoz, Mariana
Chirinos, M.
Busso, Dolores
Raffo, F. G. E.
Battistone, Maria Agustina
Blaquier, J. A.
Larrea, F.
Cuasnicu, Patricia Sara
author_role author
author2 Weigel Muñoz, Mariana
Chirinos, M.
Busso, Dolores
Raffo, F. G. E.
Battistone, Maria Agustina
Blaquier, J. A.
Larrea, F.
Cuasnicu, Patricia Sara
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv FERTILIZATION
HCRISP1
OOCYTE
SPERM
ZONA PELLUCIDA
topic FERTILIZATION
HCRISP1
OOCYTE
SPERM
ZONA PELLUCIDA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm-ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm?egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein?protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved notonly in sperm?egg fusion but also in the previous stage of sperm?ZP binding through its specific interaction with human ZP3.
Fil: Maldera, Julieta Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina. Heidelberg University. Center for Molecular Biology; Alemania
Fil: Weigel Muñoz, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Chirinos, M.. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran; México
Fil: Busso, Dolores. Pontificia Universidad Católica de Chile; Chile. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Raffo, F. G. E.. Centro Médico Fertilab; Argentina
Fil: Battistone, Maria Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Blaquier, J. A.. Centro Médico Fertilab; Argentina
Fil: Larrea, F.. Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubiran; México
Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
description Human epididymal CRISP1 (hCRISP1) associates with sperm during maturation and participates in gamete fusion through egg complementary sites. Its homology with both rodent epididymal CRISP1 and CRISP4 reported to participate in the previous stage of sperm binding to the zona pellucida (ZP), led us to further investigate the functional role of hCRISP1 by studying its involvement in human sperm-ZP interaction. Human hemizona (HZ) were inseminated with human capacitated sperm in the presence of either anti-hCRISP1 polyclonal antibody to inhibit sperm hCRISP1, or bacterially-expressed hCRISP1 (rec-hCRISP1) to block putative hCRISP1 binding sites in the ZP. Results revealed that both anti-hCRISP1 and rec-hCRISP1 produced a significant inhibition in the number of sperm bound per HZ compared with the corresponding controls. The finding that neither anti-hCRISP1 nor rec-hCRISP1 affected capacitation-associated events (i.e. sperm motility, protein tyrosine phosphorylation or acrosome reaction) supports a specific inhibition at the sperm?egg interaction level. Moreover, immunofluorescence experiments using human ZP-intact eggs revealed the presence of complementary sites for hCRISP1 in the ZP. To identify the ligand of hCRISP1 in the ZP, human recombinant proteins ZP2, ZP3 and ZP4 expressed in insect cells were co-incubated with hCRISP1 and protein?protein interaction was analyzed by ELISA. Results revealed that rec-hCRISP1 mainly interacted with ZP3 in a dose-dependent and saturable manner, supporting the specificity of this interaction. Altogether, these results indicate that hCRISP1 is a multifunctional protein involved notonly in sperm?egg fusion but also in the previous stage of sperm?ZP binding through its specific interaction with human ZP3.
publishDate 2014
dc.date.none.fl_str_mv 2014-04-01
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/11336/6291
Maldera, Julieta Antonella; Weigel Muñoz, Mariana; Chirinos, M.; Busso, Dolores; Raffo, F. G. E.; et al.; Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3; Oxford University Press; Molecular Human Reproduction; 20; 4; 1-4-2014; 341-349
1360-9947
1460-2407
url http://hdl.handle.net/11336/6291
identifier_str_mv Maldera, Julieta Antonella; Weigel Muñoz, Mariana; Chirinos, M.; Busso, Dolores; Raffo, F. G. E.; et al.; Human fertilization: epididymal hCRISP1 mediates sperm zona pellucida binding through its interaction with ZP3; Oxford University Press; Molecular Human Reproduction; 20; 4; 1-4-2014; 341-349
1360-9947
1460-2407
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/
info:eu-repo/semantics/altIdentifier/url/http://molehr.oxfordjournals.org/content/20/4/341.long
info:eu-repo/semantics/altIdentifier/doi/10.1093/molehr/gat092
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
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
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 13.070432

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