The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells

We have studied how the F-actin cytoskeleton is involved in establishing the heterogeneous intracellular Ca2+ levels ([Ca2+]i) and in the organization of the exocytotic machinery in cultured bovine chromaffin cells. Simultaneous confocal visualization of [Ca2+]i and transmitted light studies of the...

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Autores: Torregrosa-Hetland, Cristina J., Villanueva, José, Giner, Daniel, López-Font, Inmaculada, Nadal, Ángel, Expósito-Romero, Giovanna, Gil Gómez, Amparo|||0000-0002-7449-4205, González Vélez, Virginia, Segura Sala, José Javier|||0000-0002-0841-5636, Gutiérrez Pérez, Luis Miguel
Tipo de recurso: artículo
Fecha de publicación:2011
País:España
Institución:Universidad de Cantabria (UC)
Repositorio:UCrea Repositorio Abierto de la Universidad de Cantabria
Idioma:inglés
OAI Identifier:oai:repositorio.unican.es:10902/1619
Acceso en línea:http://hdl.handle.net/10902/1619
Access Level:acceso abierto
Palabra clave:Chromaffin cell
Exocytosis
F-actin cytoskeleton
Intracellular Ca2+
SNARE protein
Fluorescence microscopy
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spelling The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cellsTorregrosa-Hetland, Cristina J.Villanueva, JoséGiner, DanielLópez-Font, InmaculadaNadal, ÁngelExpósito-Romero, GiovannaGil Gómez, Amparo|||0000-0002-7449-4205González Vélez, VirginiaSegura Sala, José Javier|||0000-0002-0841-5636Gutiérrez Pérez, Luis MiguelChromaffin cellExocytosisF-actin cytoskeletonIntracellular Ca2+SNARE proteinFluorescence microscopyWe have studied how the F-actin cytoskeleton is involved in establishing the heterogeneous intracellular Ca2+ levels ([Ca2+]i) and in the organization of the exocytotic machinery in cultured bovine chromaffin cells. Simultaneous confocal visualization of [Ca2+]i and transmitted light studies of the cytoskeleton showed that, following cell stimulation, the maximal signal from the Ca2+-sensitive fluorescent dye Fluo-3 was in the empty cytosolic spaces left by cytoskeletal cages. This was mostly due to the accumulation of the dye in spaces devoid of cytoskeletal components, as shown by the use of alternative Ca2+-insensitive fluorescent cytosolic markers. In addition to affecting the distribution of such compounds in the cytosol, the cytoskeleton influenced the location of L- and P-Q-type Ca2+ channel clusters, which were associated with the borders of cytoskeletal cages in resting and stimulated cells. Indeed, syntaxin-1 and synaptotagmin-1, which are components of the secretory machinery, were present in the same location. Furthermore, granule exocytosis took place at these sites, indicating that the organization of the F-actin cytoskeletal cortex shapes the preferential sites for secretion by associating the secretory machinery with preferential sites for Ca2+ entry. The influence of this cortical organization on the propagation of [Ca2+]i can be modelled, illustrating how it serves to define rapid exocytosis.This work was supported by grants from the Spanish Ministry of Education and Culture (MEC, MICINN, BFU2005-02154/BFI and BFU2008-00731) and the Generalitat Valenciana (ACOMP06/036) to L.M.G.; the Spanish Ministry of Education and Culture (BFU2007- 67607) and AN (BFU2008-01492) to I.Q.; and the Fundación BBVA and I-MATH project C3-0136 to A.G. and J.S. C.J.T.-H. and I.L.-F. were recipients of fellowships from the MICINN (Spain). V.G-V. thanks CONACyT for its financial support of her PhD scholarship. We also acknowledge the financial support received from the CONSOLIDER programme (CSD07-00023) and CIBERDEM (Instituto de Salud Carlos III).The Company of BiologistUniversidad de Cantabria20112011-03-01journal articlehttp://purl.org/coar/resource_type/c_6501NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/articlehttp://hdl.handle.net/10902/1619Journal of Cell Science, 2011, 124 (5), 727-734.reponame:UCrea Repositorio Abierto de la Universidad de Cantabriainstname:Universidad de Cantabria (UC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.unican.es:10902/16192026-06-02T12:39:31Z
dc.title.none.fl_str_mv The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
title The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
spellingShingle The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
Torregrosa-Hetland, Cristina J.
Chromaffin cell
Exocytosis
F-actin cytoskeleton
Intracellular Ca2+
SNARE protein
Fluorescence microscopy
title_short The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
title_full The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
title_fullStr The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
title_full_unstemmed The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
title_sort The F-actin cortical network is a major factor influencing the organization of the secretory machinery in chromaffin cells
dc.creator.none.fl_str_mv Torregrosa-Hetland, Cristina J.
Villanueva, José
Giner, Daniel
López-Font, Inmaculada
Nadal, Ángel
Expósito-Romero, Giovanna
Gil Gómez, Amparo|||0000-0002-7449-4205
González Vélez, Virginia
Segura Sala, José Javier|||0000-0002-0841-5636
Gutiérrez Pérez, Luis Miguel
author Torregrosa-Hetland, Cristina J.
author_facet Torregrosa-Hetland, Cristina J.
Villanueva, José
Giner, Daniel
López-Font, Inmaculada
Nadal, Ángel
Expósito-Romero, Giovanna
Gil Gómez, Amparo|||0000-0002-7449-4205
González Vélez, Virginia
Segura Sala, José Javier|||0000-0002-0841-5636
Gutiérrez Pérez, Luis Miguel
author_role author
author2 Villanueva, José
Giner, Daniel
López-Font, Inmaculada
Nadal, Ángel
Expósito-Romero, Giovanna
Gil Gómez, Amparo|||0000-0002-7449-4205
González Vélez, Virginia
Segura Sala, José Javier|||0000-0002-0841-5636
Gutiérrez Pérez, Luis Miguel
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad de Cantabria
dc.subject.none.fl_str_mv Chromaffin cell
Exocytosis
F-actin cytoskeleton
Intracellular Ca2+
SNARE protein
Fluorescence microscopy
topic Chromaffin cell
Exocytosis
F-actin cytoskeleton
Intracellular Ca2+
SNARE protein
Fluorescence microscopy
description We have studied how the F-actin cytoskeleton is involved in establishing the heterogeneous intracellular Ca2+ levels ([Ca2+]i) and in the organization of the exocytotic machinery in cultured bovine chromaffin cells. Simultaneous confocal visualization of [Ca2+]i and transmitted light studies of the cytoskeleton showed that, following cell stimulation, the maximal signal from the Ca2+-sensitive fluorescent dye Fluo-3 was in the empty cytosolic spaces left by cytoskeletal cages. This was mostly due to the accumulation of the dye in spaces devoid of cytoskeletal components, as shown by the use of alternative Ca2+-insensitive fluorescent cytosolic markers. In addition to affecting the distribution of such compounds in the cytosol, the cytoskeleton influenced the location of L- and P-Q-type Ca2+ channel clusters, which were associated with the borders of cytoskeletal cages in resting and stimulated cells. Indeed, syntaxin-1 and synaptotagmin-1, which are components of the secretory machinery, were present in the same location. Furthermore, granule exocytosis took place at these sites, indicating that the organization of the F-actin cytoskeletal cortex shapes the preferential sites for secretion by associating the secretory machinery with preferential sites for Ca2+ entry. The influence of this cortical organization on the propagation of [Ca2+]i can be modelled, illustrating how it serves to define rapid exocytosis.
publishDate 2011
dc.date.none.fl_str_mv 2011
2011-03-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10902/1619
url http://hdl.handle.net/10902/1619
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv The Company of Biologist
publisher.none.fl_str_mv The Company of Biologist
dc.source.none.fl_str_mv Journal of Cell Science, 2011, 124 (5), 727-734.
reponame:UCrea Repositorio Abierto de la Universidad de Cantabria
instname:Universidad de Cantabria (UC)
instname_str Universidad de Cantabria (UC)
reponame_str UCrea Repositorio Abierto de la Universidad de Cantabria
collection UCrea Repositorio Abierto de la Universidad de Cantabria
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300719

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