The added value of rabies virus as a retrograde tracer when combined with dual anterograde tract-tracing

Rabies virus (RV) has widely been used as a trans-synaptic retrograde tracer to analyze chains of connected neurons. The use of antibodies directed against the viral nucleoprotein enables viral nucleocapsids to be visualized within the cell soma, as well as within the thickest main dendrites. Howeve...

Full description

Saved in:
Bibliographic Details
Published in:Journal of neuroscience methods 2010-12, Vol.194 (1), p.21-7
Main Authors: López, Iciar P, Salin, Pascal, Kachidian, Philippe, Barroso-Chinea, Pedro, Rico, Alberto J, Gómez-Bautista, Virginia, Conte-Perales, Lorena, Coulon, Patrice, Goff, Lydia Kerkerian-Le, Lanciego, José L
Format: Article
Language:English
Subjects:
Animals
Antibodies
Biotin
Cellular Biology
Cerebral Cortex
Corpus Striatum
Dextrans
Fluorescent Dyes
Image Processing, Computer-Assisted
Immunohistochemistry
Interneurons
Life Sciences
Male
Microscopy, Confocal
Neuroanatomy
Neuronal Tract-Tracers
Neurons
Phosphoproteins
Phytohemagglutinins
Rabies virus
Rats
Rats, Wistar
Synapses
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Rabies virus (RV) has widely been used as a trans-synaptic retrograde tracer to analyze chains of connected neurons. The use of antibodies directed against the viral nucleoprotein enables viral nucleocapsids to be visualized within the cell soma, as well as within the thickest main dendrites. However, through this approach it is often difficult to accurately define post-synaptic elements (thin dendrites and/or dendritic spines). This limitation can now easily been circumvented by taking advantage of antibodies directed against a soluble viral phosphoprotein that spreads throughout the cytoplasm of the infected neuron, thereby producing Golgi-like immunofluorescent labeling of first-order projection neurons that are infected with RV. Furthermore, when combined with anterograde tracers such as Phaseolus vulgaris-leucoagglutinin (PHA-L) and biotinylated dextran amine (BDA), this procedure to detect RV facilitates the accurate visualization of both the pre- and post-synaptic elements. Finally, this method of viral detection is sufficiently sensitive to detect weakly labeled second-order neurons, which can then be further characterized neurochemically. Several examples are provided to illustrate why retrograde trans-synaptic tracing using RV can be regarded as an important breakthrough in the analysis of brain circuits, providing an unprecedented level of resolution.
ISSN:0165-0270
DOI:10.1016/j.jneumeth.2010.01.015