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Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):E1082-8. doi: 10.1073/pnas.1507109113. Epub 2015 Dec 28.

Pan-neuronal imaging in roaming Caenorhabditis elegans.

Author information

1
Department of Physics, Harvard University, Cambridge, MA 02138; Center for Brain Science, Harvard University, Cambridge, MA 02138; vivek@physics.harvard.edu samuel@physics.harvard.edu.
2
Department of Physics, Harvard University, Cambridge, MA 02138; Center for Brain Science, Harvard University, Cambridge, MA 02138;
3
Department of Neurobiology, University of Massachusetts Medical School, Worcester, MA 01605;
4
Department of Physics, Nanjing University, 210093 Nanjing, China;
5
Center for Brain Science, Harvard University, Cambridge, MA 02138;
6
Division of Biological Sciences, Section of Neurobiology, University of California, San Diego, La Jolla, CA 92093;
7
Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, MA 01609;
8
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, ON, Canada M5G 1X5; Departments of Molecular Genetics and Physiology, University of Toronto, Toronto, ON, Canada M5S 1A8.

Abstract

We present an imaging system for pan-neuronal recording in crawling Caenorhabditis elegans. A spinning disk confocal microscope, modified for automated tracking of the C. elegans head ganglia, simultaneously records the activity and position of ∼80 neurons that coexpress cytoplasmic calcium indicator GCaMP6s and nuclear localized red fluorescent protein at 10 volumes per second. We developed a behavioral analysis algorithm that maps the movements of the head ganglia to the animal's posture and locomotion. Image registration and analysis software automatically assigns an index to each nucleus and calculates the corresponding calcium signal. Neurons with highly stereotyped positions can be associated with unique indexes and subsequently identified using an atlas of the worm nervous system. To test our system, we analyzed the brainwide activity patterns of moving worms subjected to thermosensory inputs. We demonstrate that our setup is able to uncover representations of sensory input and motor output of individual neurons from brainwide dynamics. Our imaging setup and analysis pipeline should facilitate mapping circuits for sensory to motor transformation in transparent behaving animals such as C. elegans and Drosophila larva.

KEYWORDS:

C. elegans; Drosophila; calcium imaging; thermotaxis

Comment in

PMID:
26711989
PMCID:
PMC4776525
DOI:
10.1073/pnas.1507109113
[Indexed for MEDLINE]
Free PMC Article

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