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Front Neural Circuits. 2018 Nov 21;12:94. doi: 10.3389/fncir.2018.00094. eCollection 2018.

A Pipeline for Volume Electron Microscopy of the Caenorhabditis elegans Nervous System.

Author information

1
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada.
2
Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
3
Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, ON, Canada.
4
Nanoscale Biomedical Imaging Facility, The Hospital for Sick Children, Peter Gilgan Centre for Research and Learning, Toronto, ON, Canada.
5
Center for Brain Science, Department of Physics, Harvard University, Cambridge, MA, United States.
6
Section of Cell and Developmental Biology, Division of Biological Sciences, University of California, San Diego, La Jolla, CA, United States.
7
Department of Physiology, University of Toronto, Toronto, ON, Canada.
8
Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada.

Abstract

The "connectome," a comprehensive wiring diagram of synaptic connectivity, is achieved through volume electron microscopy (vEM) analysis of an entire nervous system and all associated non-neuronal tissues. White et al. (1986) pioneered the fully manual reconstruction of a connectome using Caenorhabditis elegans. Recent advances in vEM allow mapping new C. elegans connectomes with increased throughput, and reduced subjectivity. Current vEM studies aim to not only fill the remaining gaps in the original connectome, but also address fundamental questions including how the connectome changes during development, the nature of individuality, sexual dimorphism, and how genetic and environmental factors regulate connectivity. Here we describe our current vEM pipeline and projected improvements for the study of the C. elegans nervous system and beyond.

KEYWORDS:

C. elegans; connectome; high-pressure freezing; nervous system; volume electron microscopy

PMID:
30524248
PMCID:
PMC6262311
DOI:
10.3389/fncir.2018.00094
[Indexed for MEDLINE]
Free PMC Article

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