Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts

Autores
Bayonés, Lucas; Guerra Fernández, María José; Hinostroza, Fernando; Báez Matus, Ximena; Vásquez Navarrete, Jacqueline; Gallo, Luciana Ines; Parra, Sergio; Martínez, Agustín D.; González Jamett, Arlek; Marengo, Fernando Diego; Cárdenas, Ana M.
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Gain-of-function mutations of dynamin-2, a mechano-GTPase that remodels membrane and actin filaments, cause centronuclear myopathy (CNM), a congenital disease that mainly affects skeletal muscle tissue. Among these mutations, the variants p.A618T and p.S619L lead to a gain of function and cause a severe neonatal phenotype. By using total internal reflection fluorescence microscopy (TIRFM) in immortalized human myoblasts expressing the pH-sensitive fluorescent protein (pHluorin) fused to the insulin-responsive aminopeptidase IRAP as a reporter of the GLUT4 vesicle trafficking, we measured single pHluorin signals to investigate how p.A618T and p.S619L mutations influence exocytosis. We show here that both dynamin-2 mutations significantly reduced the number and durations of pHluorin signals induced by 10 μM ionomycin, indicating that in addition to impairing exocytosis, they also affect the fusion pore dynamics. These mutations also disrupt the formation of actin filaments, a process that reportedly favors exocytosis. This altered exocytosis might importantly disturb the plasmalemma expression of functional proteins such as the glucose transporter GLUT4 in skeletal muscle cells, impacting the physiology of the skeletal muscle tissue and contributing to the CNM disease.
Fil: Bayonés, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Guerra Fernández, María José. Universidad de Valparaíso; Chile
Fil: Hinostroza, Fernando. Universidad Catolica de Maule; Chile
Fil: Báez Matus, Ximena. Universidad de Valparaíso; Chile
Fil: Vásquez Navarrete, Jacqueline. Universidad de Valparaíso; Chile
Fil: Gallo, Luciana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Parra, Sergio. Universidad de Valparaíso; Chile
Fil: Martínez, Agustín D.. Universidad de Valparaíso; Chile
Fil: González Jamett, Arlek. Universidad de Valparaíso; Chile
Fil: Marengo, Fernando Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Cárdenas, Ana M.. Universidad de Valparaíso; Chile
Materia
CENTRONUCLEAR MYOPATHY
DYNAMIN
DYNAMIN-2 MUTATIONS
ENDOCYTOSIS
EXOCYTOSIS
GLUT4
IRAP
PHLUORIN
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/212783
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/212783
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human MyoblastsBayonés, LucasGuerra Fernández, María JoséHinostroza, FernandoBáez Matus, XimenaVásquez Navarrete, JacquelineGallo, Luciana InesParra, SergioMartínez, Agustín D.González Jamett, ArlekMarengo, Fernando DiegoCárdenas, Ana M.CENTRONUCLEAR MYOPATHYDYNAMINDYNAMIN-2 MUTATIONSENDOCYTOSISEXOCYTOSISGLUT4IRAPPHLUORINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Gain-of-function mutations of dynamin-2, a mechano-GTPase that remodels membrane and actin filaments, cause centronuclear myopathy (CNM), a congenital disease that mainly affects skeletal muscle tissue. Among these mutations, the variants p.A618T and p.S619L lead to a gain of function and cause a severe neonatal phenotype. By using total internal reflection fluorescence microscopy (TIRFM) in immortalized human myoblasts expressing the pH-sensitive fluorescent protein (pHluorin) fused to the insulin-responsive aminopeptidase IRAP as a reporter of the GLUT4 vesicle trafficking, we measured single pHluorin signals to investigate how p.A618T and p.S619L mutations influence exocytosis. We show here that both dynamin-2 mutations significantly reduced the number and durations of pHluorin signals induced by 10 μM ionomycin, indicating that in addition to impairing exocytosis, they also affect the fusion pore dynamics. These mutations also disrupt the formation of actin filaments, a process that reportedly favors exocytosis. This altered exocytosis might importantly disturb the plasmalemma expression of functional proteins such as the glucose transporter GLUT4 in skeletal muscle cells, impacting the physiology of the skeletal muscle tissue and contributing to the CNM disease.Fil: Bayonés, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Guerra Fernández, María José. Universidad de Valparaíso; ChileFil: Hinostroza, Fernando. Universidad Catolica de Maule; ChileFil: Báez Matus, Ximena. Universidad de Valparaíso; ChileFil: Vásquez Navarrete, Jacqueline. Universidad de Valparaíso; ChileFil: Gallo, Luciana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Parra, Sergio. Universidad de Valparaíso; ChileFil: Martínez, Agustín D.. Universidad de Valparaíso; ChileFil: González Jamett, Arlek. Universidad de Valparaíso; ChileFil: Marengo, Fernando Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Cárdenas, Ana M.. Universidad de Valparaíso; ChileMDPI2022-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/212783Bayonés, Lucas; Guerra Fernández, María José; Hinostroza, Fernando; Báez Matus, Ximena; Vásquez Navarrete, Jacqueline; et al.; Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts; MDPI; International Journal of Molecular Sciences; 23; 18; 9-2022; 1-171661-65961422-0067CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1422-0067/23/18/10363info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms231810363info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:14:39Zoai:ri.conicet.gov.ar:11336/212783instacron: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 10:14:39.65CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
title Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
spellingShingle Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
Bayonés, Lucas
CENTRONUCLEAR MYOPATHY
DYNAMIN
DYNAMIN-2 MUTATIONS
ENDOCYTOSIS
EXOCYTOSIS
GLUT4
IRAP
PHLUORIN
title_short Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
title_full Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
title_fullStr Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
title_full_unstemmed Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
title_sort Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts
dc.creator.none.fl_str_mv Bayonés, Lucas
Guerra Fernández, María José
Hinostroza, Fernando
Báez Matus, Ximena
Vásquez Navarrete, Jacqueline
Gallo, Luciana Ines
Parra, Sergio
Martínez, Agustín D.
González Jamett, Arlek
Marengo, Fernando Diego
Cárdenas, Ana M.
author Bayonés, Lucas
author_facet Bayonés, Lucas
Guerra Fernández, María José
Hinostroza, Fernando
Báez Matus, Ximena
Vásquez Navarrete, Jacqueline
Gallo, Luciana Ines
Parra, Sergio
Martínez, Agustín D.
González Jamett, Arlek
Marengo, Fernando Diego
Cárdenas, Ana M.
author_role author
author2 Guerra Fernández, María José
Hinostroza, Fernando
Báez Matus, Ximena
Vásquez Navarrete, Jacqueline
Gallo, Luciana Ines
Parra, Sergio
Martínez, Agustín D.
González Jamett, Arlek
Marengo, Fernando Diego
Cárdenas, Ana M.
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CENTRONUCLEAR MYOPATHY
DYNAMIN
DYNAMIN-2 MUTATIONS
ENDOCYTOSIS
EXOCYTOSIS
GLUT4
IRAP
PHLUORIN
topic CENTRONUCLEAR MYOPATHY
DYNAMIN
DYNAMIN-2 MUTATIONS
ENDOCYTOSIS
EXOCYTOSIS
GLUT4
IRAP
PHLUORIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Gain-of-function mutations of dynamin-2, a mechano-GTPase that remodels membrane and actin filaments, cause centronuclear myopathy (CNM), a congenital disease that mainly affects skeletal muscle tissue. Among these mutations, the variants p.A618T and p.S619L lead to a gain of function and cause a severe neonatal phenotype. By using total internal reflection fluorescence microscopy (TIRFM) in immortalized human myoblasts expressing the pH-sensitive fluorescent protein (pHluorin) fused to the insulin-responsive aminopeptidase IRAP as a reporter of the GLUT4 vesicle trafficking, we measured single pHluorin signals to investigate how p.A618T and p.S619L mutations influence exocytosis. We show here that both dynamin-2 mutations significantly reduced the number and durations of pHluorin signals induced by 10 μM ionomycin, indicating that in addition to impairing exocytosis, they also affect the fusion pore dynamics. These mutations also disrupt the formation of actin filaments, a process that reportedly favors exocytosis. This altered exocytosis might importantly disturb the plasmalemma expression of functional proteins such as the glucose transporter GLUT4 in skeletal muscle cells, impacting the physiology of the skeletal muscle tissue and contributing to the CNM disease.
Fil: Bayonés, Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Guerra Fernández, María José. Universidad de Valparaíso; Chile
Fil: Hinostroza, Fernando. Universidad Catolica de Maule; Chile
Fil: Báez Matus, Ximena. Universidad de Valparaíso; Chile
Fil: Vásquez Navarrete, Jacqueline. Universidad de Valparaíso; Chile
Fil: Gallo, Luciana Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Parra, Sergio. Universidad de Valparaíso; Chile
Fil: Martínez, Agustín D.. Universidad de Valparaíso; Chile
Fil: González Jamett, Arlek. Universidad de Valparaíso; Chile
Fil: Marengo, Fernando Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Cárdenas, Ana M.. Universidad de Valparaíso; Chile
description Gain-of-function mutations of dynamin-2, a mechano-GTPase that remodels membrane and actin filaments, cause centronuclear myopathy (CNM), a congenital disease that mainly affects skeletal muscle tissue. Among these mutations, the variants p.A618T and p.S619L lead to a gain of function and cause a severe neonatal phenotype. By using total internal reflection fluorescence microscopy (TIRFM) in immortalized human myoblasts expressing the pH-sensitive fluorescent protein (pHluorin) fused to the insulin-responsive aminopeptidase IRAP as a reporter of the GLUT4 vesicle trafficking, we measured single pHluorin signals to investigate how p.A618T and p.S619L mutations influence exocytosis. We show here that both dynamin-2 mutations significantly reduced the number and durations of pHluorin signals induced by 10 μM ionomycin, indicating that in addition to impairing exocytosis, they also affect the fusion pore dynamics. These mutations also disrupt the formation of actin filaments, a process that reportedly favors exocytosis. This altered exocytosis might importantly disturb the plasmalemma expression of functional proteins such as the glucose transporter GLUT4 in skeletal muscle cells, impacting the physiology of the skeletal muscle tissue and contributing to the CNM disease.
publishDate 2022
dc.date.none.fl_str_mv 2022-09
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/11336/212783
Bayonés, Lucas; Guerra Fernández, María José; Hinostroza, Fernando; Báez Matus, Ximena; Vásquez Navarrete, Jacqueline; et al.; Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts; MDPI; International Journal of Molecular Sciences; 23; 18; 9-2022; 1-17
1661-6596
1422-0067
CONICET Digital
CONICET
url http://hdl.handle.net/11336/212783
identifier_str_mv Bayonés, Lucas; Guerra Fernández, María José; Hinostroza, Fernando; Báez Matus, Ximena; Vásquez Navarrete, Jacqueline; et al.; Gain-of-Function Dynamin-2 Mutations Linked to Centronuclear Myopathy Impair Ca2+-Induced Exocytosis in Human Myoblasts; MDPI; International Journal of Molecular Sciences; 23; 18; 9-2022; 1-17
1661-6596
1422-0067
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://www.mdpi.com/1422-0067/23/18/10363
info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms231810363
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
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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