Journal
FRONTIERS IN NEUROANATOMY
Volume 9, Issue -, Pages -Publisher
FRONTIERS RESEARCH FOUNDATION
DOI: 10.3389/fnana.7015.00060
Keywords
dendritic spines; synapses; 3D-reconstruction; electron microscopy; FIB/SEM; adult neurogenesis
Categories
Funding
- Spanish MINECO [BFU2008-03980, SAF2013-42445R, SAF2013-2010-19930, PIE13-00027, BFU2012-34963]
- CIBERNED
- Cajal Blue Brain Project, Spanish partner of the Blue Brain Project initiative from EPFL
- European Union [604102]
- ICREA Funding Source: Custom
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available