Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión

Dynamin is a GTPase required for the vesicle pinching off from the plasma membrane during the late steps of endocytosis. Growing evidences suggest that dynamin also participates duringregulated exocytosis; however its specific contribution is still elusive. Here, using bovine chromaffincells, we sho...

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Detalles Bibliográficos
Autor: González-Jamett, Arlek Marion
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2013
País:Chile
Idioma:inglés
OAI Identifier:oai:repositorio.anid.cl:10533/179881
Acceso en línea:http://creativecommons.org/licenses/by-nc-nd/3.0/cl/
https://hdl.handle.net/10533/179881
Access Level:acceso abierto
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spelling Universidad de ValparaísoGonzález-Jamett, Arlek Marion2014https://hdl.handle.net/10533/179881http://purl.org/coar/access_right/c_abf2Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatiónCárdenas-Díaz, Ana MaríaMartínez, AgustinUniversidad de ValparaísochiValparaísoGonzález-Jamett, Arlek Marion2017-03-27T22:40:57Z2022-08-17T22:18:27Z2017-03-27T22:40:57Z2022-08-17T22:18:27Z20142013Dynamin is a GTPase required for the vesicle pinching off from the plasma membrane during the late steps of endocytosis. Growing evidences suggest that dynamin also participates duringregulated exocytosis; however its specific contribution is still elusive. Here, using bovine chromaffincells, we show for the first time that the ubiquitously expressed isoform dynamin-2 controls differentstages of the exoytosis in a stimulus-dependent manner. Thus, we observed that the inhibition of dynamin-2 GTP-ase activity by the expression of a dominant negative mutant (Dyn2K44A), or theknock-down of endogenous dynamin-2 by small interfering RNA (iRNADyn2) prolonged the lifetime of the initial fusion pare, increased the catecholamine quantal release and lengthened the duration of the exocytotic events evoked by low concentration of the nicotinic agonist 1, 1-dimethyl-4-phenylpiperazinium (DMPP) (10 uM), but not during exocytotic events triggered by higher DMPP concentration (50 uM).The expression of Dyn2K44A and iRNADyn2 also dramatically disrupted cortical actin organization by displaying a punctuated pattern. Moreover, the acute inhibition of the dynamin GTP-ase activity with dynasore or dynole 34-2 completely suppressed the de novo Ca2+-dependent polymerization, resembling the effects of cytochalasin D (CytoD), an actin-disrupting drug. The treatment ofchromaffin cells with CytoD also increased the lifetime of the fusion pare, the quantal size and duration of the exocytotic events induced by 10 uM DMPP, without affecting the exocytosis induced by 50 uM DMPP. The effects of dynamin-2 disruption and cytochalasin D on the exocytosis were non-additive, suggesting that dynamin-2 and F-actin work through a common pathway controllingdifferent stages of the exocytosis.PFCHA-BecasDoctor en Ciencias Mención Neurociencia135p.PFCHA-BecasTERMINADAhttp://creativecommons.org/licenses/by-nc-nd/3.0/cl/https://hdl.handle.net/10533/179881enginstname: Conicytreponame: Repositorio Digital RI2.0instname: Conicytreponame: Repositorio Digital RI2.0handle/10533/108040info:eu-repo/grantAgreement/PFCHA-Becas/RI20info:eu-repo/semantics/dataset/hdl.handle.net/10533/93488Atribución-NoComercial-SinDerivadas 3.0 Chileinfo:eu-repo/semantics/openAccessDynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatiónTesis Doctoradoinfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersionTesisTesishttps://hdl.handle.net/10533/179881PFCHA-BecasORIGINALGONZALEZ_ARLEK_2076D.pdfapplication/pdf45782759https://repositorio.anid.cl/bitstreams/39651906-e895-46c6-9223-d838a0c739be/download6c0ffdccbfa82a29fcd0b9c0b949ea24MD51TEXTGONZALEZ_ARLEK_2076D.pdf.txtExtracted texttext/plain230893https://repositorio.anid.cl/bitstreams/4b46eee1-918e-45c3-9a6f-2d6504eb759a/downloaddbea04b7010e896fdd2b0cad22700ad7MD52THUMBNAILGONZALEZ_ARLEK_2076D.pdf.jpgIM Thumbnailimage/jpeg1926https://repositorio.anid.cl/bitstreams/7e2b95fe-3eb8-477c-a647-cfabcf57a6d0/downloadecb6e88aa2e0a8c25c6db44f27f7acb3MD5310533/179881oai:repositorio.anid.cl:10533/1798812023-07-24 04:01:51.953https://repositorio.anid.clRepositorio ANIDaletelier@anid.cl
dc.title.none.fl_str_mv Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
title Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
spellingShingle Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
González-Jamett, Arlek Marion
title_short Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
title_full Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
title_fullStr Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
title_full_unstemmed Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
title_sort Dynamin-2 regulates the late steps of exocytosis in adrenal chromaffin cells through a mechanism that involves actin polymerizatión
dc.creator.none.fl_str_mv González-Jamett, Arlek Marion
author González-Jamett, Arlek Marion
author_facet González-Jamett, Arlek Marion
author_role author
dc.contributor.advisor.none.fl_str_mv Cárdenas-Díaz, Ana María
Martínez, Agustin
dc.contributor.institution.none.fl_str_mv Universidad de Valparaíso
description Dynamin is a GTPase required for the vesicle pinching off from the plasma membrane during the late steps of endocytosis. Growing evidences suggest that dynamin also participates duringregulated exocytosis; however its specific contribution is still elusive. Here, using bovine chromaffincells, we show for the first time that the ubiquitously expressed isoform dynamin-2 controls differentstages of the exoytosis in a stimulus-dependent manner. Thus, we observed that the inhibition of dynamin-2 GTP-ase activity by the expression of a dominant negative mutant (Dyn2K44A), or theknock-down of endogenous dynamin-2 by small interfering RNA (iRNADyn2) prolonged the lifetime of the initial fusion pare, increased the catecholamine quantal release and lengthened the duration of the exocytotic events evoked by low concentration of the nicotinic agonist 1, 1-dimethyl-4-phenylpiperazinium (DMPP) (10 uM), but not during exocytotic events triggered by higher DMPP concentration (50 uM).The expression of Dyn2K44A and iRNADyn2 also dramatically disrupted cortical actin organization by displaying a punctuated pattern. Moreover, the acute inhibition of the dynamin GTP-ase activity with dynasore or dynole 34-2 completely suppressed the de novo Ca2+-dependent polymerization, resembling the effects of cytochalasin D (CytoD), an actin-disrupting drug. The treatment ofchromaffin cells with CytoD also increased the lifetime of the fusion pare, the quantal size and duration of the exocytotic events induced by 10 uM DMPP, without affecting the exocytosis induced by 50 uM DMPP. The effects of dynamin-2 disruption and cytochalasin D on the exocytosis were non-additive, suggesting that dynamin-2 and F-actin work through a common pathway controllingdifferent stages of the exocytosis.
publishDate 2013
dc.date.start.none.fl_str_mv 2013
dc.date.issued.none.fl_str_mv 2014
dc.date.accessioned.none.fl_str_mv 2017-03-27T22:40:57Z
2022-08-17T22:18:27Z
dc.date.available.none.fl_str_mv 2017-03-27T22:40:57Z
2022-08-17T22:18:27Z
dc.type.none.fl_str_mv Tesis Doctorado
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