High-Content Imaging of Immunofluorescently Labeled TRPV1-Positive Sensory Neurons

Methods Mol Biol. 2019:1987:111-124. doi: 10.1007/978-1-4939-9446-5_8.

Abstract

Studying TRP channel expressing nociceptors requires the identification of the respective subpopulations as well as the quantification of dynamic cellular events. However, the heterogeneity of sensory neurons and associated nonneuronal cells demands the analysis of large numbers of cells to reflect the distribution of entire populations. Here we report a detailed workflow how to apply high-content screening (HCS) microscopy to signaling events in TRPV1-positive neurons as well as an approach to use the selective elimination of TRPV1 positive cells from dissociated rat sensory ganglia as base for transcriptomic analysis of TRPV1-positive cells and/or as control for TRPV1 antibody specificity.

Keywords: Antibody selectivity; CGRP; CaMKIIα; HCS; High-content screening microscopy; IB4; Immunocytochemistry; Nav1.8; Nociceptive neurons; PKA; RIIβ; Sensory neurons; TRPV1.

MeSH terms

  • Animals
  • Cells, Cultured
  • Fluorescent Antibody Technique / methods
  • Male
  • Mice, Inbred C57BL
  • Microscopy / methods
  • Microscopy, Fluorescence
  • Nociceptors / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Sensory Receptor Cells / metabolism*
  • TRPV Cation Channels / immunology
  • TRPV Cation Channels / metabolism*

Substances

  • TRPV Cation Channels
  • Trpv1 protein, rat