Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation

Autores
Maldera, Julieta Antonella; Vasen, Gustavo; Ernesto, Juan Ignacio; Weigel Muñoz, Mariana; Cohen, Debora Juana; Cuasnicu, Patricia Sara
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Rat epididymal protein CRISP1 (Cysteine-Rich Secretory Protein 1) associates with sperm during maturation and participates in fertilization. Evidence indicates the existence of two populations of CRISP1 in sperm: one loosely bound, released during capacitation, and one strongly bound, that remains after this process. However, the mechanisms underlying CRISP1 binding to sperm remain mostly unknown. Considering the high concentrations of Zinc present in the epidydimis, we investigated the potential involvement of this cation in the association of CRISP1 with sperm. Caput sperm were co-incubated with epididymal fluid in the presence or absence of Zinc and binding of CRISP1 to sperm analyzed by Western blot. An increase in CRISP1 was detected in sperm exposed to Zinc but not if the cation was added with EDTA. The same results were obtained using purified CRISP1. CRISP1 association with sperm was dependent on epididymal fluid and concentrations, and incubation time. Treatment with NaCl (0.6 M) removed the in vitro-bound CRISP1 indicating that it corresponds to the loosely-bound population. Flow cytometry analysis of caput sperm exposed to biotinylated CRISP1/avidin-FITC revealed that only the cells incubated with exhibited an increase in fluorescence. When these sperm were examined by epifluorescence microscopy, a clear staining in the tail accompanied by a weaker labeling in the head was observed.Both the change in the tryptophan fluorescence emission spectra of CRISP1 when exposed to Zinc and the detection of two potential binding sites in a modeled CRISP1 three-dimensional structure, supported the interaction between CRISP1 and Zinc. Incubation of either cauda epididymal fluid or purified CRISP1 with Zinc followed by native-PAGE and Western blot revealed the presence of high molecular weight CRISP1 complexes not detected in fluids treated with EDTA. Altogether, these results suggest that CRISP1-Zinc omplexes would be involved in the association of the loosely bound population of CRISP1 with sperm during epididymal maturation.
Fil: Maldera, Julieta Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Vasen, Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Ernesto, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Weigel Muñoz, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Cohen, Debora Juana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Materia
Crisp1
Zinc
Sperm
Epididymis
Gamete Biology
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:11336/10384
Acceder
id CONICETDig_cc72d55c0f156fdf961f0319fd8394ba
oai_identifier_str oai:ri.conicet.gov.ar:11336/10384
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturationMaldera, Julieta AntonellaVasen, GustavoErnesto, Juan IgnacioWeigel Muñoz, MarianaCohen, Debora JuanaCuasnicu, Patricia SaraCrisp1ZincSpermEpididymisGamete Biologyhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Rat epididymal protein CRISP1 (Cysteine-Rich Secretory Protein 1) associates with sperm during maturation and participates in fertilization. Evidence indicates the existence of two populations of CRISP1 in sperm: one loosely bound, released during capacitation, and one strongly bound, that remains after this process. However, the mechanisms underlying CRISP1 binding to sperm remain mostly unknown. Considering the high concentrations of Zinc present in the epidydimis, we investigated the potential involvement of this cation in the association of CRISP1 with sperm. Caput sperm were co-incubated with epididymal fluid in the presence or absence of Zinc and binding of CRISP1 to sperm analyzed by Western blot. An increase in CRISP1 was detected in sperm exposed to Zinc but not if the cation was added with EDTA. The same results were obtained using purified CRISP1. CRISP1 association with sperm was dependent on epididymal fluid and concentrations, and incubation time. Treatment with NaCl (0.6 M) removed the in vitro-bound CRISP1 indicating that it corresponds to the loosely-bound population. Flow cytometry analysis of caput sperm exposed to biotinylated CRISP1/avidin-FITC revealed that only the cells incubated with exhibited an increase in fluorescence. When these sperm were examined by epifluorescence microscopy, a clear staining in the tail accompanied by a weaker labeling in the head was observed.Both the change in the tryptophan fluorescence emission spectra of CRISP1 when exposed to Zinc and the detection of two potential binding sites in a modeled CRISP1 three-dimensional structure, supported the interaction between CRISP1 and Zinc. Incubation of either cauda epididymal fluid or purified CRISP1 with Zinc followed by native-PAGE and Western blot revealed the presence of high molecular weight CRISP1 complexes not detected in fluids treated with EDTA. Altogether, these results suggest that CRISP1-Zinc omplexes would be involved in the association of the loosely bound population of CRISP1 with sperm during epididymal maturation.Fil: Maldera, Julieta Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Vasen, Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Ernesto, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Weigel Muñoz, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Cohen, Debora Juana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaSociety For The Study Of Reproduction2011-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/10384Maldera, Julieta Antonella; Vasen, Gustavo; Ernesto, Juan Ignacio; Weigel Muñoz, Mariana; Cohen, Debora Juana; et al.; Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation; Society For The Study Of Reproduction; Biology Of Reproduction; 85; 3; 9-2011; 503-5100006-33631529-7268enginfo:eu-repo/semantics/altIdentifier/url/http://www.biolreprod.org/content/85/3/503info:eu-repo/semantics/altIdentifier/url/http://dx.doi.org/10.1095/biolreprod.111.091439info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:01:21Zoai:ri.conicet.gov.ar:11336/10384instacron: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 10:01:22.686CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
title Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
spellingShingle Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
Maldera, Julieta Antonella
Crisp1
Zinc
Sperm
Epididymis
Gamete Biology
title_short Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
title_full Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
title_fullStr Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
title_full_unstemmed Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
title_sort Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation
dc.creator.none.fl_str_mv Maldera, Julieta Antonella
Vasen, Gustavo
Ernesto, Juan Ignacio
Weigel Muñoz, Mariana
Cohen, Debora Juana
Cuasnicu, Patricia Sara
author Maldera, Julieta Antonella
author_facet Maldera, Julieta Antonella
Vasen, Gustavo
Ernesto, Juan Ignacio
Weigel Muñoz, Mariana
Cohen, Debora Juana
Cuasnicu, Patricia Sara
author_role author
author2 Vasen, Gustavo
Ernesto, Juan Ignacio
Weigel Muñoz, Mariana
Cohen, Debora Juana
Cuasnicu, Patricia Sara
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Crisp1
Zinc
Sperm
Epididymis
Gamete Biology
topic Crisp1
Zinc
Sperm
Epididymis
Gamete Biology
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Rat epididymal protein CRISP1 (Cysteine-Rich Secretory Protein 1) associates with sperm during maturation and participates in fertilization. Evidence indicates the existence of two populations of CRISP1 in sperm: one loosely bound, released during capacitation, and one strongly bound, that remains after this process. However, the mechanisms underlying CRISP1 binding to sperm remain mostly unknown. Considering the high concentrations of Zinc present in the epidydimis, we investigated the potential involvement of this cation in the association of CRISP1 with sperm. Caput sperm were co-incubated with epididymal fluid in the presence or absence of Zinc and binding of CRISP1 to sperm analyzed by Western blot. An increase in CRISP1 was detected in sperm exposed to Zinc but not if the cation was added with EDTA. The same results were obtained using purified CRISP1. CRISP1 association with sperm was dependent on epididymal fluid and concentrations, and incubation time. Treatment with NaCl (0.6 M) removed the in vitro-bound CRISP1 indicating that it corresponds to the loosely-bound population. Flow cytometry analysis of caput sperm exposed to biotinylated CRISP1/avidin-FITC revealed that only the cells incubated with exhibited an increase in fluorescence. When these sperm were examined by epifluorescence microscopy, a clear staining in the tail accompanied by a weaker labeling in the head was observed.Both the change in the tryptophan fluorescence emission spectra of CRISP1 when exposed to Zinc and the detection of two potential binding sites in a modeled CRISP1 three-dimensional structure, supported the interaction between CRISP1 and Zinc. Incubation of either cauda epididymal fluid or purified CRISP1 with Zinc followed by native-PAGE and Western blot revealed the presence of high molecular weight CRISP1 complexes not detected in fluids treated with EDTA. Altogether, these results suggest that CRISP1-Zinc omplexes would be involved in the association of the loosely bound population of CRISP1 with sperm during epididymal maturation.
Fil: Maldera, Julieta Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Vasen, Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Ernesto, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Weigel Muñoz, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Cohen, Debora Juana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina
description Rat epididymal protein CRISP1 (Cysteine-Rich Secretory Protein 1) associates with sperm during maturation and participates in fertilization. Evidence indicates the existence of two populations of CRISP1 in sperm: one loosely bound, released during capacitation, and one strongly bound, that remains after this process. However, the mechanisms underlying CRISP1 binding to sperm remain mostly unknown. Considering the high concentrations of Zinc present in the epidydimis, we investigated the potential involvement of this cation in the association of CRISP1 with sperm. Caput sperm were co-incubated with epididymal fluid in the presence or absence of Zinc and binding of CRISP1 to sperm analyzed by Western blot. An increase in CRISP1 was detected in sperm exposed to Zinc but not if the cation was added with EDTA. The same results were obtained using purified CRISP1. CRISP1 association with sperm was dependent on epididymal fluid and concentrations, and incubation time. Treatment with NaCl (0.6 M) removed the in vitro-bound CRISP1 indicating that it corresponds to the loosely-bound population. Flow cytometry analysis of caput sperm exposed to biotinylated CRISP1/avidin-FITC revealed that only the cells incubated with exhibited an increase in fluorescence. When these sperm were examined by epifluorescence microscopy, a clear staining in the tail accompanied by a weaker labeling in the head was observed.Both the change in the tryptophan fluorescence emission spectra of CRISP1 when exposed to Zinc and the detection of two potential binding sites in a modeled CRISP1 three-dimensional structure, supported the interaction between CRISP1 and Zinc. Incubation of either cauda epididymal fluid or purified CRISP1 with Zinc followed by native-PAGE and Western blot revealed the presence of high molecular weight CRISP1 complexes not detected in fluids treated with EDTA. Altogether, these results suggest that CRISP1-Zinc omplexes would be involved in the association of the loosely bound population of CRISP1 with sperm during epididymal maturation.
publishDate 2011
dc.date.none.fl_str_mv 2011-09
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/11336/10384
Maldera, Julieta Antonella; Vasen, Gustavo; Ernesto, Juan Ignacio; Weigel Muñoz, Mariana; Cohen, Debora Juana; et al.; Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation; Society For The Study Of Reproduction; Biology Of Reproduction; 85; 3; 9-2011; 503-510
0006-3363
1529-7268
url http://hdl.handle.net/11336/10384
identifier_str_mv Maldera, Julieta Antonella; Vasen, Gustavo; Ernesto, Juan Ignacio; Weigel Muñoz, Mariana; Cohen, Debora Juana; et al.; Evidence for the involvement of zinc in the association of crisp1 with sperm during epididymal maturation; Society For The Study Of Reproduction; Biology Of Reproduction; 85; 3; 9-2011; 503-510
0006-3363
1529-7268
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.biolreprod.org/content/85/3/503
info:eu-repo/semantics/altIdentifier/url/http://dx.doi.org/10.1095/biolreprod.111.091439
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 Society For The Study Of Reproduction
publisher.none.fl_str_mv Society For The Study Of Reproduction
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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