Calcium imaging of neuronal circuits in vivo using a circuit-tracing pseudorabies virus.

Publication Year
2010

Type

Journal Article
Abstract

Pseudorabies virus (PRV) is a neuroinvasive virus of the herpes family that has a broad host range but does not infect higher-order primates. PRV characteristically travels along chains of synaptically connected neurons and has been used extensively for elucidating neural circuits in the peripheral and central nervous system in vivo. The recombinant virus PRV369 is an attenuated retrograde tracer that encodes G-CaMP2, a fluorescent calcium sensor protein that is stable at physiological pH and mammalian temperature. This protocol describes the use of PRV369 to express G-CaMP2 in a neuronal circuit and to monitor its activity in a living animal, specifically in the submandibular ganglia (SMG), the peripheral parasympathetic ganglia that innervate the salivary glands. The procedure describes the delivery of PRV369 to the glands and shows how SMG neurons can then be imaged post-inoculation to explore connectivity and activity.

Journal
Cold Spring Harb Protoc
Volume
2010
Issue
4
Pages
pdb.prot5410
Date Published
04/2010
ISSN Number
1559-6095
Alternate Journal
Cold Spring Harb Protoc
PMID
20360364