Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells

Autores
Takarada, Takeshi; Kou, Miki; Nakamichi, Noritaka; Ogura, Masato; Ito, Yuma; Fukumori, Ryo; Kokubo, Hiroshi; Acosta, Gabriela Beatriz; Hinoi, Eiichi; Yoneda, Yukio
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background:We have previously shown marked upregulation of the mRNA and corresponding protein for the cellular motor molecule myosin VI (Myo6) after an extremely traumatic stress experience, along with a delayed decrease in 5-bromo-2′-deoxyuridine incorporation in the murine hippocampus, a brain structure believed to undergo adult neurogenesis. In this study, we investigated the role of Myo6 in both proliferation and differentiation in pluripotent P19 cells by using stable transfection and RNA interference techniques.Methodology/Principal Findings:Stable overexpression of Myo6 not only led to significant inhibition of the reducing activity of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and the size of clustered aggregates in P19 cells, but also resulted in selectively decreased mRNA expression of the repressor type proneural gene Hes5 without affecting the expression of neuronal and astroglial marker proteins. In P19 cells transfected with Myo6 siRNA, by contrast, a significant increase was found in the size of aggregate and MTT reduction along with increased Sox2 protein levels, in addition to marked depletion of the endogenous Myo6 protein. In C6 glioma cells, however, introduction of Myo6 siRNA induced a drastic decrease in endogenous Myo6 protein levels without significantly affecting MTT reduction. The Ca2+ ionophore A23187 drastically increased the luciferase activity in P19 cells transfected with a Myo6 promoter reporter plasmid, but not in HEK293, Neuro2A and C6 glioma cells transfected with the same reporter.Conclusions/Significance:These results suggest that Myo6 may play a predominant pivotal role in the mechanism underlying proliferation without affecting differentiation to progeny lineages in pluripotent P19 cells.
Fil: Takarada, Takeshi. Kanazawa University; Japón
Fil: Kou, Miki. Kanazawa University; Japón
Fil: Nakamichi, Noritaka. Kanazawa University; Japón
Fil: Ogura, Masato. Kanazawa University; Japón
Fil: Ito, Yuma. Kanazawa University; Japón
Fil: Fukumori, Ryo. Kanazawa University; Japón
Fil: Kokubo, Hiroshi. Kanazawa University; Japón
Fil: Acosta, Gabriela Beatriz. Kanazawa University; Japón. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; Argentina
Fil: Hinoi, Eiichi. Kanazawa University; Japón
Fil: Yoneda, Yukio. Kanazawa University; Japón
Materia
molecule myosin VI (Myo6)
proliferation and differentiation in pluripotent P19 cells
mRNA expression
Ca2+ ionophore A23187
HEK293, Neuro2A and C6 glioma cells
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:11336/484
Acceder
id CONICETDig_980bec04f80177939baba34f02d4ae59
oai_identifier_str oai:ri.conicet.gov.ar:11336/484
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 CellsTakarada, TakeshiKou, MikiNakamichi, NoritakaOgura, MasatoIto, YumaFukumori, RyoKokubo, HiroshiAcosta, Gabriela BeatrizHinoi, EiichiYoneda, Yukiomolecule myosin VI (Myo6)proliferation and differentiation in pluripotent P19 cellsmRNA expressionCa2+ ionophore A23187HEK293, Neuro2A and C6 glioma cellshttps://purl.org/becyt/ford/3.3https://purl.org/becyt/ford/3Background:We have previously shown marked upregulation of the mRNA and corresponding protein for the cellular motor molecule myosin VI (Myo6) after an extremely traumatic stress experience, along with a delayed decrease in 5-bromo-2′-deoxyuridine incorporation in the murine hippocampus, a brain structure believed to undergo adult neurogenesis. In this study, we investigated the role of Myo6 in both proliferation and differentiation in pluripotent P19 cells by using stable transfection and RNA interference techniques.Methodology/Principal Findings:Stable overexpression of Myo6 not only led to significant inhibition of the reducing activity of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and the size of clustered aggregates in P19 cells, but also resulted in selectively decreased mRNA expression of the repressor type proneural gene Hes5 without affecting the expression of neuronal and astroglial marker proteins. In P19 cells transfected with Myo6 siRNA, by contrast, a significant increase was found in the size of aggregate and MTT reduction along with increased Sox2 protein levels, in addition to marked depletion of the endogenous Myo6 protein. In C6 glioma cells, however, introduction of Myo6 siRNA induced a drastic decrease in endogenous Myo6 protein levels without significantly affecting MTT reduction. The Ca2+ ionophore A23187 drastically increased the luciferase activity in P19 cells transfected with a Myo6 promoter reporter plasmid, but not in HEK293, Neuro2A and C6 glioma cells transfected with the same reporter.Conclusions/Significance:These results suggest that Myo6 may play a predominant pivotal role in the mechanism underlying proliferation without affecting differentiation to progeny lineages in pluripotent P19 cells.Fil: Takarada, Takeshi. Kanazawa University; JapónFil: Kou, Miki. Kanazawa University; JapónFil: Nakamichi, Noritaka. Kanazawa University; JapónFil: Ogura, Masato. Kanazawa University; JapónFil: Ito, Yuma. Kanazawa University; JapónFil: Fukumori, Ryo. Kanazawa University; JapónFil: Kokubo, Hiroshi. Kanazawa University; JapónFil: Acosta, Gabriela Beatriz. Kanazawa University; Japón. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; ArgentinaFil: Hinoi, Eiichi. Kanazawa University; JapónFil: Yoneda, Yukio. Kanazawa University; JapónPublic Library of Science2013-05-17info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/484Takarada, Takeshi; Kou, Miki; Nakamichi, Noritaka; Ogura, Masato; Ito, Yuma; et al.; Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells; Public Library of Science; Plos One; 8; 5; 17-5-2013; 1-11; e639471932-6203CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0063947info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063947info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-04-23T14:56:55Zoai:ri.conicet.gov.ar:11336/484instacron: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:34982026-04-23 14:56:55.886CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
title Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
spellingShingle Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
Takarada, Takeshi
molecule myosin VI (Myo6)
proliferation and differentiation in pluripotent P19 cells
mRNA expression
Ca2+ ionophore A23187
HEK293, Neuro2A and C6 glioma cells
title_short Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
title_full Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
title_fullStr Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
title_full_unstemmed Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
title_sort Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells
dc.creator.none.fl_str_mv Takarada, Takeshi
Kou, Miki
Nakamichi, Noritaka
Ogura, Masato
Ito, Yuma
Fukumori, Ryo
Kokubo, Hiroshi
Acosta, Gabriela Beatriz
Hinoi, Eiichi
Yoneda, Yukio
author Takarada, Takeshi
author_facet Takarada, Takeshi
Kou, Miki
Nakamichi, Noritaka
Ogura, Masato
Ito, Yuma
Fukumori, Ryo
Kokubo, Hiroshi
Acosta, Gabriela Beatriz
Hinoi, Eiichi
Yoneda, Yukio
author_role author
author2 Kou, Miki
Nakamichi, Noritaka
Ogura, Masato
Ito, Yuma
Fukumori, Ryo
Kokubo, Hiroshi
Acosta, Gabriela Beatriz
Hinoi, Eiichi
Yoneda, Yukio
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv molecule myosin VI (Myo6)
proliferation and differentiation in pluripotent P19 cells
mRNA expression
Ca2+ ionophore A23187
HEK293, Neuro2A and C6 glioma cells
topic molecule myosin VI (Myo6)
proliferation and differentiation in pluripotent P19 cells
mRNA expression
Ca2+ ionophore A23187
HEK293, Neuro2A and C6 glioma cells
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.3
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Background:We have previously shown marked upregulation of the mRNA and corresponding protein for the cellular motor molecule myosin VI (Myo6) after an extremely traumatic stress experience, along with a delayed decrease in 5-bromo-2′-deoxyuridine incorporation in the murine hippocampus, a brain structure believed to undergo adult neurogenesis. In this study, we investigated the role of Myo6 in both proliferation and differentiation in pluripotent P19 cells by using stable transfection and RNA interference techniques.Methodology/Principal Findings:Stable overexpression of Myo6 not only led to significant inhibition of the reducing activity of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and the size of clustered aggregates in P19 cells, but also resulted in selectively decreased mRNA expression of the repressor type proneural gene Hes5 without affecting the expression of neuronal and astroglial marker proteins. In P19 cells transfected with Myo6 siRNA, by contrast, a significant increase was found in the size of aggregate and MTT reduction along with increased Sox2 protein levels, in addition to marked depletion of the endogenous Myo6 protein. In C6 glioma cells, however, introduction of Myo6 siRNA induced a drastic decrease in endogenous Myo6 protein levels without significantly affecting MTT reduction. The Ca2+ ionophore A23187 drastically increased the luciferase activity in P19 cells transfected with a Myo6 promoter reporter plasmid, but not in HEK293, Neuro2A and C6 glioma cells transfected with the same reporter.Conclusions/Significance:These results suggest that Myo6 may play a predominant pivotal role in the mechanism underlying proliferation without affecting differentiation to progeny lineages in pluripotent P19 cells.
Fil: Takarada, Takeshi. Kanazawa University; Japón
Fil: Kou, Miki. Kanazawa University; Japón
Fil: Nakamichi, Noritaka. Kanazawa University; Japón
Fil: Ogura, Masato. Kanazawa University; Japón
Fil: Ito, Yuma. Kanazawa University; Japón
Fil: Fukumori, Ryo. Kanazawa University; Japón
Fil: Kokubo, Hiroshi. Kanazawa University; Japón
Fil: Acosta, Gabriela Beatriz. Kanazawa University; Japón. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Farmacológicas. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones Farmacológicas; Argentina
Fil: Hinoi, Eiichi. Kanazawa University; Japón
Fil: Yoneda, Yukio. Kanazawa University; Japón
description Background:We have previously shown marked upregulation of the mRNA and corresponding protein for the cellular motor molecule myosin VI (Myo6) after an extremely traumatic stress experience, along with a delayed decrease in 5-bromo-2′-deoxyuridine incorporation in the murine hippocampus, a brain structure believed to undergo adult neurogenesis. In this study, we investigated the role of Myo6 in both proliferation and differentiation in pluripotent P19 cells by using stable transfection and RNA interference techniques.Methodology/Principal Findings:Stable overexpression of Myo6 not only led to significant inhibition of the reducing activity of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and the size of clustered aggregates in P19 cells, but also resulted in selectively decreased mRNA expression of the repressor type proneural gene Hes5 without affecting the expression of neuronal and astroglial marker proteins. In P19 cells transfected with Myo6 siRNA, by contrast, a significant increase was found in the size of aggregate and MTT reduction along with increased Sox2 protein levels, in addition to marked depletion of the endogenous Myo6 protein. In C6 glioma cells, however, introduction of Myo6 siRNA induced a drastic decrease in endogenous Myo6 protein levels without significantly affecting MTT reduction. The Ca2+ ionophore A23187 drastically increased the luciferase activity in P19 cells transfected with a Myo6 promoter reporter plasmid, but not in HEK293, Neuro2A and C6 glioma cells transfected with the same reporter.Conclusions/Significance:These results suggest that Myo6 may play a predominant pivotal role in the mechanism underlying proliferation without affecting differentiation to progeny lineages in pluripotent P19 cells.
publishDate 2013
dc.date.none.fl_str_mv 2013-05-17
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/11336/484
Takarada, Takeshi; Kou, Miki; Nakamichi, Noritaka; Ogura, Masato; Ito, Yuma; et al.; Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells; Public Library of Science; Plos One; 8; 5; 17-5-2013; 1-11; e63947
1932-6203
CONICET Digital
CONICET
url http://hdl.handle.net/11336/484
identifier_str_mv Takarada, Takeshi; Kou, Miki; Nakamichi, Noritaka; Ogura, Masato; Ito, Yuma; et al.; Myosin VI Reduces Proliferation, but Not Differentiation, in Pluripotent P19 Cells; Public Library of Science; Plos One; 8; 5; 17-5-2013; 1-11; e63947
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.0063947
info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063947
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
_version_ 1863366074499596288
score 13.05261

Publicaciones similares