FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons

The fine analysis of synaptic contacts is usually performed using transmission electron microscopy (TEM) and its combination with neuronal labeling techniques. However, the complex 3D architecture of neuronal samples calls for their reconstruction from serial sections. Here we show that focused ion...

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Autores: Bosch, Carles, Martínez García, Albert, Masachs Janoher, Núria, Teixeira, Catia Marlene, Fernaud, Isabel, Ulloa Darquea, Fausto Alexander, Pérez Martínez, Mª Esther, Lois, Carlos, Comella i Carnicé, Joan Xavier, 1963-, DeFelipe, Javier, Merchán Pérez, Angel, Soriano García, Eduardo
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2015
País:España
Institución:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/138540
Acceso en línea:https://hdl.handle.net/2445/138540
Access Level:acceso abierto
Palabra clave:Neurones
Sinapsi
Neurons
Synapses
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spelling FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neuronsBosch, CarlesMartínez García, AlbertMasachs Janoher, NúriaTeixeira, Catia MarleneFernaud, IsabelUlloa Darquea, Fausto AlexanderPérez Martínez, Mª EstherLois, CarlosComella i Carnicé, Joan Xavier, 1963-DeFelipe, JavierMerchán Pérez, AngelSoriano García, EduardoNeuronesSinapsiNeuronsSynapsesThe fine analysis of synaptic contacts is usually performed using transmission electron microscopy (TEM) and its combination with neuronal labeling techniques. However, the complex 3D architecture of neuronal samples calls for their reconstruction from serial sections. Here we show that focused ion beam/scanning electron microscopy (FIB/SEM) allows efficient, complete, and automatic 3D reconstruction of identified dendrites, including their spines and synapses, from GFP/DAB-labeled neurons, with a resolution comparable to that of TEM. We applied this technology to analyze the synaptogenesis of labeled adult-generated granule cells (GCs) in mice. 3D reconstruction of dendritic spines in GCs aged 3-4 and 8-9 weeks revealed two different stages of dendritic spine development and unexpected features of synapse formation, including vacant and branched dendritic spines and presynaptic terminals establishing synapses with up to 10 dendritic spines. Given the reliability, efficiency, and high resolution of FIB/SEM technology and the wide use of DAB in conventional EM, we consider FIB/SEM fundamental for the detailed characterization of identified synaptic contacts in neurons in a high-throughput manner.Frontiers Media2015info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://hdl.handle.net/2445/138540Articles publicats en revistes (Biologia Cel·lular, Fisiologia i Immunologia)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésReproducció del document publicat a: https://doi.org/10.3389/fnana.2015.00060Frontiers In Neuroanatomy, 2015, vol. 9, p. 60https://doi.org/10.3389/fnana.2015.00060info:eu-repo/grantAgreement/EC/FP7/604102cc-by (c) Bosch, Carles et al., 2015http://creativecommons.org/licenses/by/3.0/esinfo:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/1385402026-05-27T06:46:51Z
dc.title.none.fl_str_mv FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
title FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
spellingShingle FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
Bosch, Carles
Neurones
Sinapsi
Neurons
Synapses
title_short FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
title_full FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
title_fullStr FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
title_full_unstemmed FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
title_sort FIB/SEM technology allows highthroughput 3D reconstruction of dendritic spines and synapses in GFP-traced adult-generated neurons
dc.creator.none.fl_str_mv Bosch, Carles
Martínez García, Albert
Masachs Janoher, Núria
Teixeira, Catia Marlene
Fernaud, Isabel
Ulloa Darquea, Fausto Alexander
Pérez Martínez, Mª Esther
Lois, Carlos
Comella i Carnicé, Joan Xavier, 1963-
DeFelipe, Javier
Merchán Pérez, Angel
Soriano García, Eduardo
author Bosch, Carles
author_facet Bosch, Carles
Martínez García, Albert
Masachs Janoher, Núria
Teixeira, Catia Marlene
Fernaud, Isabel
Ulloa Darquea, Fausto Alexander
Pérez Martínez, Mª Esther
Lois, Carlos
Comella i Carnicé, Joan Xavier, 1963-
DeFelipe, Javier
Merchán Pérez, Angel
Soriano García, Eduardo
author_role author
author2 Martínez García, Albert
Masachs Janoher, Núria
Teixeira, Catia Marlene
Fernaud, Isabel
Ulloa Darquea, Fausto Alexander
Pérez Martínez, Mª Esther
Lois, Carlos
Comella i Carnicé, Joan Xavier, 1963-
DeFelipe, Javier
Merchán Pérez, Angel
Soriano García, Eduardo
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Neurones
Sinapsi
Neurons
Synapses
topic Neurones
Sinapsi
Neurons
Synapses
description The fine analysis of synaptic contacts is usually performed using transmission electron microscopy (TEM) and its combination with neuronal labeling techniques. However, the complex 3D architecture of neuronal samples calls for their reconstruction from serial sections. Here we show that focused ion beam/scanning electron microscopy (FIB/SEM) allows efficient, complete, and automatic 3D reconstruction of identified dendrites, including their spines and synapses, from GFP/DAB-labeled neurons, with a resolution comparable to that of TEM. We applied this technology to analyze the synaptogenesis of labeled adult-generated granule cells (GCs) in mice. 3D reconstruction of dendritic spines in GCs aged 3-4 and 8-9 weeks revealed two different stages of dendritic spine development and unexpected features of synapse formation, including vacant and branched dendritic spines and presynaptic terminals establishing synapses with up to 10 dendritic spines. Given the reliability, efficiency, and high resolution of FIB/SEM technology and the wide use of DAB in conventional EM, we consider FIB/SEM fundamental for the detailed characterization of identified synaptic contacts in neurons in a high-throughput manner.
publishDate 2015
dc.date.none.fl_str_mv 2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/138540
url https://hdl.handle.net/2445/138540
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.3389/fnana.2015.00060
Frontiers In Neuroanatomy, 2015, vol. 9, p. 60
https://doi.org/10.3389/fnana.2015.00060
info:eu-repo/grantAgreement/EC/FP7/604102
dc.rights.none.fl_str_mv cc-by (c) Bosch, Carles et al., 2015
http://creativecommons.org/licenses/by/3.0/es
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by (c) Bosch, Carles et al., 2015
http://creativecommons.org/licenses/by/3.0/es
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv Articles publicats en revistes (Biologia Cel·lular, Fisiologia i Immunologia)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,198674

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