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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Detalles Bibliográficos
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
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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