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...

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Detalles Bibliográficos
Autores: Bosch, Carles, Martínez, Albert, Masachs, Nuria, Teixeira, Cátia M., Fernaud, Isabel, Ulloa, Fausto, Pérez-Martínez, Esther, Lois, Carlos, Comella i Carnicé, Joan Xavier|||0000-0002-6218-0786, DeFelipe, Javier, Merchán-Pérez, Angel, Soriano García, Eduardo
Tipo de recurso: artículo
Fecha de publicación:2015
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:185345
Acceso en línea:https://ddd.uab.cat/record/185345
https://dx.doi.org/urn:doi:10.3389/fnana.2015.00060
Access Level:acceso abierto
Palabra clave:Dendritic spines
Synapses
3D-reconstruction
Electron microscopy
FIB/SEM
Adult neurogenesis
Descripción
Sumario: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.

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