# Protocol for the precise activation of intradental neurons in mice via electrical stimulation

## TL;DR

This paper introduces a non-invasive method to activate and study tooth neurons in mice using electrical stimulation without damaging the teeth.

## Contribution

A novel protocol for precise electrical stimulation of intradental neurons in mice with single-molar resolution.

## Key findings

- A custom stereotaxic stage and retractor system enables access to molars in anesthetized mice.
- Direct current pulses can selectively activate intradental neurons while monitoring induced current.
- The protocol supports neural imaging to study sensory responses and circuits in tooth neurons.

## Abstract

Intradental neurons encode tooth sensation, yet identifying them without sacrificing tooth structural integrity remains challenging. Here, we present a protocol for the non-invasive electrical stimulation of intradental neurons with single-molar tooth resolution in anesthetized mice. We describe details for head fixation to access molars. We then provide steps to deliver direct current (DC) pulses to an individual molar while simultaneously monitoring induced current. The protocol can be combined with neural imaging to identify and characterize intradental sensory responses and circuits.

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

•Fabricating a custom mouse stereotaxic stage and retractors for molar access•Assembly of an electrical stimulator setup to target individual molars•Procedures for selective intradental neuron activation in anesthetized mice

Fabricating a custom mouse stereotaxic stage and retractors for molar access

Assembly of an electrical stimulator setup to target individual molars

Procedures for selective intradental neuron activation in anesthetized mice

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

Intradental neurons encode tooth sensation, yet identifying them without sacrificing tooth structural integrity remains challenging. Here, we present a protocol for the non-invasive electrical stimulation of intradental neurons with single-molar tooth resolution in anesthetized mice. We describe details for head fixation to access molars. We then provide steps to deliver direct current (DC) pulses to an individual molar while simultaneously monitoring induced current. The protocol can be combined with neural imaging to identify and characterize intradental sensory responses and circuits.