Virus-Mediated Gene Transfer Into the Mouse Dorsal Cochlear Nucleus for Light-Induced Auditory Stimulation

Abstract

Source: Kozin, E. D., et.al. Direct Visualization of the Murine Dorsal Cochlear Nucleus for Optogenetic Stimulation of the Auditory Pathway. J. Vis. Exp. (2015)

This video demonstrates virus-mediated gene transfer into the mouse dorsal cochlear nucleus (DCN) to enable light-induced auditory stimulation. A viral vector encoding opsin, a light-sensitive ion channel, is injected into the DCN following a craniotomy and cerebellar aspiration for direct access. The expressed opsins allow neurons to be activated by light, providing a model for studying auditory pathway responses using optogenetics.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Cerebellar Aspiration

1. Using a 5 French suction, aspirate the lateral-most portion of the left cerebellum overlying the DCN until the DCN is visual (Figure 1, Right Panel). Remove approximately 1/4 to 1/3 of the left cerebellum. The main landmark adjacent to the DCN is the ampulla of the superior semicircular canal.
   NOTE: Aspiration of the cerebellum is the key step of the protocol. Directed aspiration of the cerebellum is ....

Results

Figure 1: Partial cerebellar aspiration demonstrates access to the dorsal cochlear nucleus. Left Panel: After the .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Stereotaxic holder	Stoelting	51500
5 French suction	Symmetry Surgical	2777914
Dental Points	Henry Schein	100-8170
Bone rongeur	Fine Surgical Tools	16020-14
10 µl Hamilton syringe	Hamilton	7633-01
34 gauge, needle	Hamilton	207434
Rongeurs	Fine Surgical Tools	16021-14