Department of Neurobiology; Lois Lab
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.
3D-reconstruction, FIB/SEM, adult neurogenesis, dendritic spines, electron microscopy, synapses
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Copyright © 2015 Bosch, Martínez, Masachs, Teixeira, Fernaud, Ulloa, Pérez-Martínez, Lois, Comella, DeFelipe, Merchán-Pérez and Soriano. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
DOI of Published Version
Front Neuroanat. 2015 May 21;9:60. doi: 10.3389/fnana.2015.00060. eCollection 2015. Link to article on publisher's site
Frontiers in neuroanatomy
Bosch, Carles; Martinez, Albert; Masachs, Nuria; Teixeira, Catia M.; Fernaud, Isabel; Ulloa, Fausto; Perez-Martinez, Esther; Lois, Carlos; Comella, Joan X.; DeFelipe, Javier; Merchan-Perez, Angel; and Soriano, Eduardo, "FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult-generated neurons" (2015). Open Access Articles. 2731.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.