Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones

Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at t...
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Autor*in:	Olivier Thoumine [verfasserIn] Matthieu Lagardère Mikael Garcia Amélie Argento Cécile Leduc Jean-Baptiste Sibarita

Format:	Artikel
Sprache:	Englisch

Erschienen:	2015

Rechteinformationen:	Nutzungsrecht: © COPYRIGHT 2015 National Academy of Sciences

Schlagwörter:	DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics

Übergeordnetes Werk:	Enthalten in: Proceedings of the National Academy of Sciences of the United States of America - Washington, DC : NAS, 1877, 112(2015), 22, Seite 6997
Übergeordnetes Werk:	volume:112 ; year:2015 ; number:22 ; pages:6997

Links:	Volltext Link aufrufen Link aufrufen Link aufrufen

DOI / URN:	10.1073/pnas.1423455112

Katalog-ID:	OLC197027154X

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520			\|a Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration.
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650		4	\|a DNA Primers - genetics
650		4	\|a Catenins - metabolism
650		4	\|a Growth Cones - metabolism
650		4	\|a Cell Movement - physiology
650		4	\|a Multiprotein Complexes - metabolism
650		4	\|a Hippocampus - cytology
650		4	\|a Actins - metabolism
650		4	\|a Embryo, Mammalian - cytology
650		4	\|a Cadherins - metabolism
650		4	\|a Protein-protein interactions
650		4	\|a Observations
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650		4	\|a Neural circuitry
650		4	\|a Physiological aspects
650		4	\|a Substrates
650		4	\|a Cytoskeleton
650		4	\|a Proteins
650		4	\|a Fluorescence
650		4	\|a Computer simulation
650		4	\|a Kinetics
700	0		\|a Matthieu Lagardère \|4 oth
700	0		\|a Mikael Garcia \|4 oth
700	0		\|a Amélie Argento \|4 oth
700	0		\|a Cécile Leduc \|4 oth
700	0		\|a Jean-Baptiste Sibarita \|4 oth
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allfields	10.1073/pnas.1423455112 doi PQ20160211 (DE-627)OLC197027154X (DE-599)GBVOLC197027154X (PRQ)g2105-333a44e1f1887f564c02cd015e2280779ccc878b5270fc9cd6ad279b4f954b953 (KEY)0583363920150000112002206997twotieredcouplingbetweenflowingactinandimmobilized DE-627 ger DE-627 rakwb eng 500 DNB 570 AVZ LING fid BIODIV fid Olivier Thoumine verfasserin aut Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration. Nutzungsrecht: © COPYRIGHT 2015 National Academy of Sciences DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics Matthieu Lagardère oth Mikael Garcia oth Amélie Argento oth Cécile Leduc oth Jean-Baptiste Sibarita oth Enthalten in Proceedings of the National Academy of Sciences of the United States of America Washington, DC : NAS, 1877 112(2015), 22, Seite 6997 (DE-627)129505269 (DE-600)209104-5 (DE-576)014909189 0027-8424 nnns volume:112 year:2015 number:22 pages:6997 http://dx.doi.org/10.1073/pnas.1423455112 Volltext http://www.pnas.org/content/112/22/6997.abstract http://www.ncbi.nlm.nih.gov/pubmed/26038554 http://search.proquest.com/docview/1688663658 GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-LING FID-BIODIV SSG-OLC-PHY SSG-OLC-CHE SSG-OLC-MAT SSG-OLC-FOR SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT SSG-OPC-FOR GBV_ILN_40 GBV_ILN_59 AR 112 2015 22 6997
spelling	10.1073/pnas.1423455112 doi PQ20160211 (DE-627)OLC197027154X (DE-599)GBVOLC197027154X (PRQ)g2105-333a44e1f1887f564c02cd015e2280779ccc878b5270fc9cd6ad279b4f954b953 (KEY)0583363920150000112002206997twotieredcouplingbetweenflowingactinandimmobilized DE-627 ger DE-627 rakwb eng 500 DNB 570 AVZ LING fid BIODIV fid Olivier Thoumine verfasserin aut Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration. Nutzungsrecht: © COPYRIGHT 2015 National Academy of Sciences DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics Matthieu Lagardère oth Mikael Garcia oth Amélie Argento oth Cécile Leduc oth Jean-Baptiste Sibarita oth Enthalten in Proceedings of the National Academy of Sciences of the United States of America Washington, DC : NAS, 1877 112(2015), 22, Seite 6997 (DE-627)129505269 (DE-600)209104-5 (DE-576)014909189 0027-8424 nnns volume:112 year:2015 number:22 pages:6997 http://dx.doi.org/10.1073/pnas.1423455112 Volltext http://www.pnas.org/content/112/22/6997.abstract http://www.ncbi.nlm.nih.gov/pubmed/26038554 http://search.proquest.com/docview/1688663658 GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-LING FID-BIODIV SSG-OLC-PHY SSG-OLC-CHE SSG-OLC-MAT SSG-OLC-FOR SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT SSG-OPC-FOR GBV_ILN_40 GBV_ILN_59 AR 112 2015 22 6997
allfields_unstemmed	10.1073/pnas.1423455112 doi PQ20160211 (DE-627)OLC197027154X (DE-599)GBVOLC197027154X (PRQ)g2105-333a44e1f1887f564c02cd015e2280779ccc878b5270fc9cd6ad279b4f954b953 (KEY)0583363920150000112002206997twotieredcouplingbetweenflowingactinandimmobilized DE-627 ger DE-627 rakwb eng 500 DNB 570 AVZ LING fid BIODIV fid Olivier Thoumine verfasserin aut Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration. Nutzungsrecht: © COPYRIGHT 2015 National Academy of Sciences DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics Matthieu Lagardère oth Mikael Garcia oth Amélie Argento oth Cécile Leduc oth Jean-Baptiste Sibarita oth Enthalten in Proceedings of the National Academy of Sciences of the United States of America Washington, DC : NAS, 1877 112(2015), 22, Seite 6997 (DE-627)129505269 (DE-600)209104-5 (DE-576)014909189 0027-8424 nnns volume:112 year:2015 number:22 pages:6997 http://dx.doi.org/10.1073/pnas.1423455112 Volltext http://www.pnas.org/content/112/22/6997.abstract http://www.ncbi.nlm.nih.gov/pubmed/26038554 http://search.proquest.com/docview/1688663658 GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-LING FID-BIODIV SSG-OLC-PHY SSG-OLC-CHE SSG-OLC-MAT SSG-OLC-FOR SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT SSG-OPC-FOR GBV_ILN_40 GBV_ILN_59 AR 112 2015 22 6997
allfieldsGer	10.1073/pnas.1423455112 doi PQ20160211 (DE-627)OLC197027154X (DE-599)GBVOLC197027154X (PRQ)g2105-333a44e1f1887f564c02cd015e2280779ccc878b5270fc9cd6ad279b4f954b953 (KEY)0583363920150000112002206997twotieredcouplingbetweenflowingactinandimmobilized DE-627 ger DE-627 rakwb eng 500 DNB 570 AVZ LING fid BIODIV fid Olivier Thoumine verfasserin aut Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration. Nutzungsrecht: © COPYRIGHT 2015 National Academy of Sciences DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics Matthieu Lagardère oth Mikael Garcia oth Amélie Argento oth Cécile Leduc oth Jean-Baptiste Sibarita oth Enthalten in Proceedings of the National Academy of Sciences of the United States of America Washington, DC : NAS, 1877 112(2015), 22, Seite 6997 (DE-627)129505269 (DE-600)209104-5 (DE-576)014909189 0027-8424 nnns volume:112 year:2015 number:22 pages:6997 http://dx.doi.org/10.1073/pnas.1423455112 Volltext http://www.pnas.org/content/112/22/6997.abstract http://www.ncbi.nlm.nih.gov/pubmed/26038554 http://search.proquest.com/docview/1688663658 GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-LING FID-BIODIV SSG-OLC-PHY SSG-OLC-CHE SSG-OLC-MAT SSG-OLC-FOR SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT SSG-OPC-FOR GBV_ILN_40 GBV_ILN_59 AR 112 2015 22 6997
allfieldsSound	10.1073/pnas.1423455112 doi PQ20160211 (DE-627)OLC197027154X (DE-599)GBVOLC197027154X (PRQ)g2105-333a44e1f1887f564c02cd015e2280779ccc878b5270fc9cd6ad279b4f954b953 (KEY)0583363920150000112002206997twotieredcouplingbetweenflowingactinandimmobilized DE-627 ger DE-627 rakwb eng 500 DNB 570 AVZ LING fid BIODIV fid Olivier Thoumine verfasserin aut Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration. Nutzungsrecht: © COPYRIGHT 2015 National Academy of Sciences DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics Matthieu Lagardère oth Mikael Garcia oth Amélie Argento oth Cécile Leduc oth Jean-Baptiste Sibarita oth Enthalten in Proceedings of the National Academy of Sciences of the United States of America Washington, DC : NAS, 1877 112(2015), 22, Seite 6997 (DE-627)129505269 (DE-600)209104-5 (DE-576)014909189 0027-8424 nnns volume:112 year:2015 number:22 pages:6997 http://dx.doi.org/10.1073/pnas.1423455112 Volltext http://www.pnas.org/content/112/22/6997.abstract http://www.ncbi.nlm.nih.gov/pubmed/26038554 http://search.proquest.com/docview/1688663658 GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-LING FID-BIODIV SSG-OLC-PHY SSG-OLC-CHE SSG-OLC-MAT SSG-OLC-FOR SSG-OLC-PHA SSG-OLC-DE-84 SSG-OPC-MAT SSG-OPC-FOR GBV_ILN_40 GBV_ILN_59 AR 112 2015 22 6997
language	English
source	Enthalten in Proceedings of the National Academy of Sciences of the United States of America 112(2015), 22, Seite 6997 volume:112 year:2015 number:22 pages:6997
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topic_facet	DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics
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author	Olivier Thoumine
spellingShingle	Olivier Thoumine ddc 500 ddc 570 fid LING fid BIODIV misc DNA Primers - genetics misc Catenins - metabolism misc Growth Cones - metabolism misc Cell Movement - physiology misc Multiprotein Complexes - metabolism misc Hippocampus - cytology misc Actins - metabolism misc Embryo, Mammalian - cytology misc Cadherins - metabolism misc Protein-protein interactions misc Observations misc Actin misc Cadherins misc Neural circuitry misc Physiological aspects misc Substrates misc Cytoskeleton misc Proteins misc Fluorescence misc Computer simulation misc Kinetics Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones
authorStr	Olivier Thoumine
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topic_title	500 DNB 570 AVZ LING fid BIODIV fid Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones DNA Primers - genetics Catenins - metabolism Growth Cones - metabolism Cell Movement - physiology Multiprotein Complexes - metabolism Hippocampus - cytology Actins - metabolism Embryo, Mammalian - cytology Cadherins - metabolism Protein-protein interactions Observations Actin Cadherins Neural circuitry Physiological aspects Substrates Cytoskeleton Proteins Fluorescence Computer simulation Kinetics
topic	ddc 500 ddc 570 fid LING fid BIODIV misc DNA Primers - genetics misc Catenins - metabolism misc Growth Cones - metabolism misc Cell Movement - physiology misc Multiprotein Complexes - metabolism misc Hippocampus - cytology misc Actins - metabolism misc Embryo, Mammalian - cytology misc Cadherins - metabolism misc Protein-protein interactions misc Observations misc Actin misc Cadherins misc Neural circuitry misc Physiological aspects misc Substrates misc Cytoskeleton misc Proteins misc Fluorescence misc Computer simulation misc Kinetics
topic_unstemmed	ddc 500 ddc 570 fid LING fid BIODIV misc DNA Primers - genetics misc Catenins - metabolism misc Growth Cones - metabolism misc Cell Movement - physiology misc Multiprotein Complexes - metabolism misc Hippocampus - cytology misc Actins - metabolism misc Embryo, Mammalian - cytology misc Cadherins - metabolism misc Protein-protein interactions misc Observations misc Actin misc Cadherins misc Neural circuitry misc Physiological aspects misc Substrates misc Cytoskeleton misc Proteins misc Fluorescence misc Computer simulation misc Kinetics
topic_browse	ddc 500 ddc 570 fid LING fid BIODIV misc DNA Primers - genetics misc Catenins - metabolism misc Growth Cones - metabolism misc Cell Movement - physiology misc Multiprotein Complexes - metabolism misc Hippocampus - cytology misc Actins - metabolism misc Embryo, Mammalian - cytology misc Cadherins - metabolism misc Protein-protein interactions misc Observations misc Actin misc Cadherins misc Neural circuitry misc Physiological aspects misc Substrates misc Cytoskeleton misc Proteins misc Fluorescence misc Computer simulation misc Kinetics
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title	Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones
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title_full	Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones
author_sort	Olivier Thoumine
journal	Proceedings of the National Academy of Sciences of the United States of America
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title_sort	two-tiered coupling between flowing actin and immobilized n-cadherin/catenin complexes in neuronal growth cones
title_auth	Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones
abstract	Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration.
abstractGer	Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration.
abstract_unstemmed	Neuronal growth cones move forward by dynamically connecting actin-based motility to substrate adhesion, but the mechanisms at the individual molecular level remain unclear. We cultured primary neurons on N-cadherin-coated micropatterned substrates, and imaged adhesion and cytoskeletal proteins at the ventral surface of growth cones using single particle tracking combined to photoactivated localization microscopy (sptPALM). We demonstrate transient interactions in the second time scale between flowing actin filaments and immobilized N-cadherin/catenin complexes, translating into a local reduction of the actin retrograde flow. Normal actin flow on micropatterns was rescued by expression of a dominant negative N-cadherin construct competing for the coupling between actin and endogenous N-cadherin. Fluorescence recovery after photobleaching (FRAP) experiments confirmed the differential kinetics of actin and N-cadherin, and further revealed a 20% actin population confined at N-cadherin micropatterns, contributing to local actin accumulation. Computer simulations with relevant kinetic parameters modeled N-cadherin and actin turnover well, validating this mechanism. Such a combination of short- and long-lived interactions between the motile actin network and spatially restricted adhesive complexes represents a two-tiered clutch mechanism likely to sustain dynamic environment sensing and provide the force necessary for growth cone migration.
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container_issue	22
title_short	Two-tiered coupling between flowing actin and immobilized N-cadherin/catenin complexes in neuronal growth cones
url	http://dx.doi.org/10.1073/pnas.1423455112 http://www.pnas.org/content/112/22/6997.abstract http://www.ncbi.nlm.nih.gov/pubmed/26038554 http://search.proquest.com/docview/1688663658
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author2	Matthieu Lagardère Mikael Garcia Amélie Argento Cécile Leduc Jean-Baptiste Sibarita
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up_date	2024-07-03T14:34:16.790Z
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