Protocol for intra-nerve AAV injection and dorsal root potential recording for optogenetic modulation of the peripheral sensory nerve activity

TL;DR

This paper provides a detailed protocol for using optogenetics to study peripheral sensory nerve activity in rats.

Contribution

The paper introduces a novel method for intra-nerve AAV injection and dorsal root potential recording in optogenetic studies.

Findings

AAV injection enables gene transduction in dorsal root ganglion neurons via retrograde transport.

Optical and electrical stimulation can be used to record dorsal root potentials in vivo.

The protocol allows for acute evaluation of optogenetic effects on peripheral sensory nerves.

Abstract

Optogenetic modulation of peripheral sensory nerve activity holds great potential for revealing the mechanisms underlying sensory disorders, with important implications for developing therapeutic strategies. Here, we present a protocol for applying optogenetic techniques to peripheral sensory nerves using an adeno-associated virus (AAV) vector. We describe the procedure for gene transduction into dorsal root ganglion neurons via retrograde transport following intra-nerve AAV injection. We then outline a terminal, acute electrophysiological experiment to evaluate optogenetic effects at the dorsal root level.

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

•Instructions for gene transduction via viral injection into the rat sciatic nerve•Steps for preparing an in vivo dorsal root potential recording at the lumbar level•Guidance on recording dorsal root potentials using electrical and optical stimulation

Instructions for gene transduction via viral injection into the rat sciatic nerve

Steps for preparing an in vivo dorsal root potential recording at the lumbar level

Guidance on recording dorsal root potentials using electrical and optical stimulation

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

Optogenetic modulation of peripheral sensory nerve activity holds great potential for revealing the mechanisms underlying sensory disorders, with important implications for developing therapeutic strategies. Here, we present a protocol for applying optogenetic techniques to peripheral sensory nerves using an adeno-associated virus (AAV) vector. We describe the procedure for gene transduction into dorsal root ganglion neurons via retrograde transport following intra-nerve AAV injection. We then outline a terminal, acute electrophysiological experiment to evaluate optogenetic effects at the dorsal root level.

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