# Protocol for targeted peripheral adeno-associated viral vector transduction of the intradental neurons in mice

## TL;DR

This paper describes a surgical method to deliver a virus to specific neurons in mouse teeth to study their connections.

## Contribution

A novel protocol for targeted adeno-associated virus transduction of intradental neurons in mice using a silk fibroin additive.

## Key findings

- AAV-i.d. successfully labels intradental neurons and their projections.
- The protocol includes precise steps for mouse positioning and drilling depth.
- A silk fibroin additive stabilizes the AAV on the tooth surface.

## Abstract

Here, we present the methodology for surgical preparation of the mouse molar teeth and subsequent adeno-associated virus (AAV) transduction of intradental neurons within the tooth (AAV-i.d.). We describe steps for mouse positioning, microscope setup, drilling to expose pulp horns, and applying the AAV-silk mixture. We then detail procedures for composite placement to cover the preparation and trigeminal ganglion (TG) screening to visualize the surface and assess and verify AAV labeling. AAV-i.d. labels intradental neurons and traces their associated neuronal projections.

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

•Workflow for novel-targeted intradental neuron transduction in the trigeminal ganglion•Description of mouse positioning for either maxillary or mandibular molar access•Stabilization of AAV on the tooth preparation using a silk fibroin additive•Guidance on drilling depth, AAV-mixture application, and labeling variability

Workflow for novel-targeted intradental neuron transduction in the trigeminal ganglion

Description of mouse positioning for either maxillary or mandibular molar access

Stabilization of AAV on the tooth preparation using a silk fibroin additive

Guidance on drilling depth, AAV-mixture application, and labeling variability

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

Here, we present the methodology for surgical preparation of the mouse molar teeth and subsequent adeno-associated virus (AAV) transduction of intradental neurons within the tooth (AAV-i.d.). We describe steps for mouse positioning, microscope setup, drilling to expose pulp horns, and applying the AAV-silk mixture. We then detail procedures for composite placement to cover the preparation and trigeminal ganglion (TG) screening to visualize the surface and assess and verify AAV labeling. AAV-i.d. labels intradental neurons and traces their associated neuronal projections.