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PLoS One. 2014 Nov 5;9(11):e112102. doi: 10.1371/journal.pone.0112102. eCollection 2014.

Isolation of specific neurons from C. elegans larvae for gene expression profiling.

Author information

1
Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee, United States of America.
2
Program in Neuroscience, Vanderbilt University, Nashville, Tennessee, United States of America.
3
Department of Genome Sciences, University of Washington School of Medicine, Seattle, Washington, United States of America.
4
Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee, United States of America; Program in Neuroscience, Vanderbilt University, Nashville, Tennessee, United States of America.

Abstract

BACKGROUND:

The simple and well-described structure of the C. elegans nervous system offers an unprecedented opportunity to identify the genetic programs that define the connectivity and function of individual neurons and their circuits. A correspondingly precise gene expression map of C. elegans neurons would facilitate the application of genetic methods toward this goal. Here we describe a powerful new approach, SeqCeL (RNA-Seq of C. elegans cells) for producing gene expression profiles of specific larval C. elegans neurons.

METHODS AND RESULTS:

We have exploited available GFP reporter lines for FACS isolation of specific larval C. elegans neurons for RNA-Seq analysis. Our analysis showed that diverse classes of neurons are accessible to this approach. To demonstrate the applicability of this strategy to rare neuron types, we generated RNA-Seq profiles of the NSM serotonergic neurons that occur as a single bilateral pair of cells in the C. elegans pharynx. These data detected >1,000 NSM enriched transcripts, including the majority of previously known NSM-expressed genes.

SIGNIFICANCE:

This work offers a simple and robust protocol for expression profiling studies of post-embryonic C. elegans neurons and thus provides an important new method for identifying candidate genes for key roles in neuron-specific development and function.

PMID:
25372608
PMCID:
PMC4221280
DOI:
10.1371/journal.pone.0112102
[Indexed for MEDLINE]
Free PMC Article

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