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 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.
Liu et al. (Sat,) studied this question.