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

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

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Caenorhabditis elegans* / cytology
  • Caenorhabditis elegans* / metabolism
  • Cell Separation / methods
  • Gene Expression Profiling*
  • Gene Expression Regulation / physiology
  • Neurons* / cytology
  • Neurons* / metabolism

Associated data

  • SRA/SRX139566
  • SRA/SRX139591
  • SRA/SRX139602
  • SRA/SRX145444
  • SRA/SRX145445
  • SRA/SRX145447
  • SRA/SRX190368