Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes

Autores
Mayorga, Luis Segundo; Altamirano, Karina Noel; Zanni Ruiz, Emilia; Pavarotti, Martin Alejandro
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Capacitation is not a well‐defined process, required for the acrosome reaction triggered by physiological stimuli. In vitro, capacitation is achieved by sperm incubation in artificial media supplemented with HCO3ˉ, Ca2+, and albumin. The role of capacitation in the membrane fusion machinery required for acrosomal exocytosis is not well known. SNAREs proteins are fundamental for intracellular membrane fusion and acrosomal exocytosis. We have previously shown that in capacitated sperm, the fusion machinery is maintained in an inactive state until the acrosome reaction is initiated. In particular, SNARE proteins are assembled in neurotoxin‐resistant complexes.Objective:This work aimed to study the dynamic changes of SNARE complexes during capacitation.Materials and Methods: The light chain of tetanus and botulinum neurotoxin has been widely used to study the configuration of SNARE proteins. For this purpose, we developed a recombinant light chain of tetanus neurotoxin linked to a polyarginine peptide. This membrane permeant protein was able to cleave cytosolic VAMP2 (a SNARE protein required for acrosome reaction) when present in a monomeric configuration.Results: The results show that the VAMP2 is cleaved by the membrane permeant tetanus neurotoxin in non‐capacitated sperm, indicating that, before capacitation, SNAREs are not assembled in stable toxin‐resistant complexes. However, 2 h of incubation in a capacitation medium containing albumin was sufficient to render VAMP2 insensitive to the toxin. Discussion: We conclude that during capacitation, the SNARE proteins become engaged in stable fully assembled cis SNARE complexes. This step is likely essential to prevent untimely activation of the membrane fusion machinery.ConclusionWe propose that capacitation promotes the stabilization of the membrane fusion machinery required for acrosomal exocytosis in preparation for the stimulus‐triggered acrosome reaction.
Fil: Mayorga, Luis Segundo. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Altamirano, Karina Noel. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Zanni Ruiz, Emilia. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo; Argentina
Fil: Pavarotti, Martin Alejandro. Universidad Nacional de Cuyo; Argentina. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Materia
SPERM
CAPACITATION
SNARE
NEUROTOXIN
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:11336/110386
Acceder
id CONICETDig_be11393680a0b88ecf8d4394eaa7a968
oai_identifier_str oai:ri.conicet.gov.ar:11336/110386
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexesMayorga, Luis SegundoAltamirano, Karina NoelZanni Ruiz, EmiliaPavarotti, Martin AlejandroSPERMCAPACITATIONSNARENEUROTOXINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Capacitation is not a well‐defined process, required for the acrosome reaction triggered by physiological stimuli. In vitro, capacitation is achieved by sperm incubation in artificial media supplemented with HCO3ˉ, Ca2+, and albumin. The role of capacitation in the membrane fusion machinery required for acrosomal exocytosis is not well known. SNAREs proteins are fundamental for intracellular membrane fusion and acrosomal exocytosis. We have previously shown that in capacitated sperm, the fusion machinery is maintained in an inactive state until the acrosome reaction is initiated. In particular, SNARE proteins are assembled in neurotoxin‐resistant complexes.Objective:This work aimed to study the dynamic changes of SNARE complexes during capacitation.Materials and Methods: The light chain of tetanus and botulinum neurotoxin has been widely used to study the configuration of SNARE proteins. For this purpose, we developed a recombinant light chain of tetanus neurotoxin linked to a polyarginine peptide. This membrane permeant protein was able to cleave cytosolic VAMP2 (a SNARE protein required for acrosome reaction) when present in a monomeric configuration.Results: The results show that the VAMP2 is cleaved by the membrane permeant tetanus neurotoxin in non‐capacitated sperm, indicating that, before capacitation, SNAREs are not assembled in stable toxin‐resistant complexes. However, 2 h of incubation in a capacitation medium containing albumin was sufficient to render VAMP2 insensitive to the toxin. Discussion: We conclude that during capacitation, the SNARE proteins become engaged in stable fully assembled cis SNARE complexes. This step is likely essential to prevent untimely activation of the membrane fusion machinery.ConclusionWe propose that capacitation promotes the stabilization of the membrane fusion machinery required for acrosomal exocytosis in preparation for the stimulus‐triggered acrosome reaction.Fil: Mayorga, Luis Segundo. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Altamirano, Karina Noel. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Zanni Ruiz, Emilia. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo; ArgentinaFil: Pavarotti, Martin Alejandro. Universidad Nacional de Cuyo; Argentina. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaJohn Wiley and Sons Inc.2020-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/110386Mayorga, Luis Segundo; Altamirano, Karina Noel; Zanni Ruiz, Emilia; Pavarotti, Martin Alejandro; Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes; John Wiley and Sons Inc.; Andrology; 8; 2; 3-2020; 442-4492047-2919CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1111/andr.12706info:eu-repo/semantics/altIdentifier/doi/10.1111/andr.12706info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:02:11Zoai:ri.conicet.gov.ar:11336/110386instacron: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 10:02:12.009CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
title Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
spellingShingle Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
Mayorga, Luis Segundo
SPERM
CAPACITATION
SNARE
NEUROTOXIN
title_short Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
title_full Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
title_fullStr Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
title_full_unstemmed Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
title_sort Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes
dc.creator.none.fl_str_mv Mayorga, Luis Segundo
Altamirano, Karina Noel
Zanni Ruiz, Emilia
Pavarotti, Martin Alejandro
author Mayorga, Luis Segundo
author_facet Mayorga, Luis Segundo
Altamirano, Karina Noel
Zanni Ruiz, Emilia
Pavarotti, Martin Alejandro
author_role author
author2 Altamirano, Karina Noel
Zanni Ruiz, Emilia
Pavarotti, Martin Alejandro
author2_role author
author
author
dc.subject.none.fl_str_mv SPERM
CAPACITATION
SNARE
NEUROTOXIN
topic SPERM
CAPACITATION
SNARE
NEUROTOXIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Capacitation is not a well‐defined process, required for the acrosome reaction triggered by physiological stimuli. In vitro, capacitation is achieved by sperm incubation in artificial media supplemented with HCO3ˉ, Ca2+, and albumin. The role of capacitation in the membrane fusion machinery required for acrosomal exocytosis is not well known. SNAREs proteins are fundamental for intracellular membrane fusion and acrosomal exocytosis. We have previously shown that in capacitated sperm, the fusion machinery is maintained in an inactive state until the acrosome reaction is initiated. In particular, SNARE proteins are assembled in neurotoxin‐resistant complexes.Objective:This work aimed to study the dynamic changes of SNARE complexes during capacitation.Materials and Methods: The light chain of tetanus and botulinum neurotoxin has been widely used to study the configuration of SNARE proteins. For this purpose, we developed a recombinant light chain of tetanus neurotoxin linked to a polyarginine peptide. This membrane permeant protein was able to cleave cytosolic VAMP2 (a SNARE protein required for acrosome reaction) when present in a monomeric configuration.Results: The results show that the VAMP2 is cleaved by the membrane permeant tetanus neurotoxin in non‐capacitated sperm, indicating that, before capacitation, SNAREs are not assembled in stable toxin‐resistant complexes. However, 2 h of incubation in a capacitation medium containing albumin was sufficient to render VAMP2 insensitive to the toxin. Discussion: We conclude that during capacitation, the SNARE proteins become engaged in stable fully assembled cis SNARE complexes. This step is likely essential to prevent untimely activation of the membrane fusion machinery.ConclusionWe propose that capacitation promotes the stabilization of the membrane fusion machinery required for acrosomal exocytosis in preparation for the stimulus‐triggered acrosome reaction.
Fil: Mayorga, Luis Segundo. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Altamirano, Karina Noel. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
Fil: Zanni Ruiz, Emilia. Universidad Nacional de Cuyo. Facultad de Ciencias Exactas y Naturales; Argentina. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina. Universidad Nacional de Cuyo; Argentina
Fil: Pavarotti, Martin Alejandro. Universidad Nacional de Cuyo; Argentina. 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 Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentina
description Capacitation is not a well‐defined process, required for the acrosome reaction triggered by physiological stimuli. In vitro, capacitation is achieved by sperm incubation in artificial media supplemented with HCO3ˉ, Ca2+, and albumin. The role of capacitation in the membrane fusion machinery required for acrosomal exocytosis is not well known. SNAREs proteins are fundamental for intracellular membrane fusion and acrosomal exocytosis. We have previously shown that in capacitated sperm, the fusion machinery is maintained in an inactive state until the acrosome reaction is initiated. In particular, SNARE proteins are assembled in neurotoxin‐resistant complexes.Objective:This work aimed to study the dynamic changes of SNARE complexes during capacitation.Materials and Methods: The light chain of tetanus and botulinum neurotoxin has been widely used to study the configuration of SNARE proteins. For this purpose, we developed a recombinant light chain of tetanus neurotoxin linked to a polyarginine peptide. This membrane permeant protein was able to cleave cytosolic VAMP2 (a SNARE protein required for acrosome reaction) when present in a monomeric configuration.Results: The results show that the VAMP2 is cleaved by the membrane permeant tetanus neurotoxin in non‐capacitated sperm, indicating that, before capacitation, SNAREs are not assembled in stable toxin‐resistant complexes. However, 2 h of incubation in a capacitation medium containing albumin was sufficient to render VAMP2 insensitive to the toxin. Discussion: We conclude that during capacitation, the SNARE proteins become engaged in stable fully assembled cis SNARE complexes. This step is likely essential to prevent untimely activation of the membrane fusion machinery.ConclusionWe propose that capacitation promotes the stabilization of the membrane fusion machinery required for acrosomal exocytosis in preparation for the stimulus‐triggered acrosome reaction.
publishDate 2020
dc.date.none.fl_str_mv 2020-03
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/11336/110386
Mayorga, Luis Segundo; Altamirano, Karina Noel; Zanni Ruiz, Emilia; Pavarotti, Martin Alejandro; Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes; John Wiley and Sons Inc.; Andrology; 8; 2; 3-2020; 442-449
2047-2919
CONICET Digital
CONICET
url http://hdl.handle.net/11336/110386
identifier_str_mv Mayorga, Luis Segundo; Altamirano, Karina Noel; Zanni Ruiz, Emilia; Pavarotti, Martin Alejandro; Human sperm capacitation is necessary for SNARE assembly in neurotoxin‐resistant complexes; John Wiley and Sons Inc.; Andrology; 8; 2; 3-2020; 442-449
2047-2919
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1111/andr.12706
info:eu-repo/semantics/altIdentifier/doi/10.1111/andr.12706
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 John Wiley and Sons Inc.
publisher.none.fl_str_mv John Wiley and Sons Inc.
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
_version_ 1842269743058255872
score 13.13397

Publicaciones similares