FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled 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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2015-05-01
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| Series: | Frontiers in Neuroanatomy |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fnana.2015.00060/full |
| _version_ | 1819137572655333376 |
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| author | Carles eBosch Carles eBosch Carles eBosch Albert eMartínez Núria eMasachs Núria eMasachs Cátia M Teixeira Cátia M Teixeira Isabel eFernaud Isabel eFernaud Isabel eFernaud Fausto eUlloa Fausto eUlloa Esther ePérez Esther ePérez Carlos eLois Joan X. Comella Joan X. Comella Joan X. Comella Javier eDeFelipe Javier eDeFelipe Javier eDeFelipe Angel eMerchan-Perez Angel eMerchan-Perez Angel eMerchan-Perez Eduardo eSoriano Eduardo eSoriano Eduardo eSoriano |
| author_facet | Carles eBosch Carles eBosch Carles eBosch Albert eMartínez Núria eMasachs Núria eMasachs Cátia M Teixeira Cátia M Teixeira Isabel eFernaud Isabel eFernaud Isabel eFernaud Fausto eUlloa Fausto eUlloa Esther ePérez Esther ePérez Carlos eLois Joan X. Comella Joan X. Comella Joan X. Comella Javier eDeFelipe Javier eDeFelipe Javier eDeFelipe Angel eMerchan-Perez Angel eMerchan-Perez Angel eMerchan-Perez Eduardo eSoriano Eduardo eSoriano Eduardo eSoriano |
| author_sort | Carles eBosch |
| collection | DOAJ |
| 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 spines in GCs aged 3–4 and 8–9 weeks revealed two different stages of spine development and unexpected features of synapse formation, including vacant and branched spines and presynaptic terminals establishing synapses with up to 10 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. |
| first_indexed | 2024-12-22T10:53:01Z |
| format | Article |
| id | doaj.art-548590a8322047acb8d30846a92cb115 |
| institution | Directory Open Access Journal |
| issn | 1662-5129 |
| language | English |
| last_indexed | 2024-12-22T10:53:01Z |
| publishDate | 2015-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Neuroanatomy |
| spelling | doaj.art-548590a8322047acb8d30846a92cb1152022-12-21T18:28:43ZengFrontiers Media S.A.Frontiers in Neuroanatomy1662-51292015-05-01910.3389/fnana.2015.00060129749FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neuronsCarles eBosch0Carles eBosch1Carles eBosch2Albert eMartínez3Núria eMasachs4Núria eMasachs5Cátia M Teixeira6Cátia M Teixeira7Isabel eFernaud8Isabel eFernaud9Isabel eFernaud10Fausto eUlloa11Fausto eUlloa12Esther ePérez13Esther ePérez14Carlos eLois15Joan X. Comella16Joan X. Comella17Joan X. Comella18Javier eDeFelipe19Javier eDeFelipe20Javier eDeFelipe21Angel eMerchan-Perez22Angel eMerchan-Perez23Angel eMerchan-Perez24Eduardo eSoriano25Eduardo eSoriano26Eduardo eSoriano27University of BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIInstitut de Recerca de l’Hospital Universitari de la Vall d’Hebron (VHIR)University of BarcelonaUniversity of BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIUniversity of BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIICentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIUniversidad Politécnica de MadridInstituto Cajal (CSIC)University of BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIUniversity of BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIUniversity of Massachusetts Medical SchoolCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIInstitut de Recerca de l’Hospital Universitari de la Vall d’Hebron (VHIR)Universitat Autònoma de BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIUniversidad Politécnica de MadridInstituto Cajal (CSIC)Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIUniversidad Politécnica de MadridUniversidad Politécnica de MadridUniversity of BarcelonaCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ISCIIIInstitut de Recerca de l’Hospital Universitari de la Vall d’Hebron (VHIR)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 spines in GCs aged 3–4 and 8–9 weeks revealed two different stages of spine development and unexpected features of synapse formation, including vacant and branched spines and presynaptic terminals establishing synapses with up to 10 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.http://journal.frontiersin.org/Journal/10.3389/fnana.2015.00060/fullDendritic SpinesSynapsesadult neurogenesis3D-reconstructionElectron microscopyFIB/SEM |
| spellingShingle | Carles eBosch Carles eBosch Carles eBosch Albert eMartínez Núria eMasachs Núria eMasachs Cátia M Teixeira Cátia M Teixeira Isabel eFernaud Isabel eFernaud Isabel eFernaud Fausto eUlloa Fausto eUlloa Esther ePérez Esther ePérez Carlos eLois Joan X. Comella Joan X. Comella Joan X. Comella Javier eDeFelipe Javier eDeFelipe Javier eDeFelipe Angel eMerchan-Perez Angel eMerchan-Perez Angel eMerchan-Perez Eduardo eSoriano Eduardo eSoriano Eduardo eSoriano FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons Frontiers in Neuroanatomy Dendritic Spines Synapses adult neurogenesis 3D-reconstruction Electron microscopy FIB/SEM |
| title | FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons |
| title_full | FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons |
| title_fullStr | FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons |
| title_full_unstemmed | FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons |
| title_short | FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons |
| title_sort | fib sem technology and high throughput 3d reconstruction of dendritic spines and synapses in gfp labeled adult generated neurons |
| topic | Dendritic Spines Synapses adult neurogenesis 3D-reconstruction Electron microscopy FIB/SEM |
| url | http://journal.frontiersin.org/Journal/10.3389/fnana.2015.00060/full |
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