# Protocol to isolate dorsal root ganglion neurons from embryonic rats by immunopanning and characterize them using RNAscope and immunofluorescence

## TL;DR

This paper provides a detailed protocol for isolating and studying dorsal root ganglion neurons from rat embryos using immunopanning and molecular techniques.

## Contribution

The novel contribution is a comprehensive protocol for purifying DRG neurons and quantifying mRNA and proteins in isolated neurons.

## Key findings

- A protocol for isolating DRG neurons from E15 rat embryos using immunopanning is described.
- Methods for mRNA and protein quantification using RNAscope and immunofluorescence are detailed.
- The protocol enables study of sensory neurons in cultures with minimal contamination from other cell types.

## Abstract

Dorsal root ganglion (DRG) neurons innervate the periphery of the body to transmit somatosensory information, including touch, pain, and temperature. Here, we present a protocol for isolating rat DRG neurons using an immunopanning technique. We describe steps for dissecting DRGs from embryonic day (E)15 rat embryos, triturating ganglionic cells, and purifying DRG neurons. We also detail procedures for mRNA or protein quantification using RNAscope and immunofluorescence. This protocol has potential applications for studying DRG neurons during development and disease.

For complete details on the use and execution of this protocol, please refer to Kantarci et al.1

•Utilizing immunopanning to isolate dorsal root ganglion neurons from rat embryos•Study of sensory neurons in cultures largely devoid of glia or other cell types•Detection of proteins of interest via immunofluorescence in isolated neurons•Guidance for using RNAscope to quantify target mRNA molecules expressed in neurons

Utilizing immunopanning to isolate dorsal root ganglion neurons from rat embryos

Study of sensory neurons in cultures largely devoid of glia or other cell types

Detection of proteins of interest via immunofluorescence in isolated neurons

Guidance for using RNAscope to quantify target mRNA molecules expressed in neurons

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Dorsal root ganglion (DRG) neurons innervate the periphery of the body to transmit somatosensory information, including touch, pain, and temperature. Here, we present a protocol for isolating rat DRG neurons using an immunopanning technique. We describe steps for dissecting DRGs from embryonic day (E)15 rat embryos, triturating ganglionic cells, and purifying DRG neurons. We also detail procedures for mRNA or protein quantification using RNAscope and immunofluorescence. This protocol has potential applications for studying DRG neurons during development and disease.