Recombinant Fluorescent Rabies Virus Vectors for Tracing Neurons and Synaptic Connections | Zendy

Nadin Hagendorf | Zendy; KarlKlaus Conzelmann | Zendy

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Recombinant Fluorescent Rabies Virus Vectors for Tracing Neurons and Synaptic Connections

Author(s) -

Nadin Hagendorf,

KarlKlaus Conzelmann

Publication year - 2015

Publication title -

cold spring harbor protocols

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.674

H-Index - 51

eISSN - 1940-3402

pISSN - 1559-6095

DOI - 10.1101/pdb.top089391

Subject(s) - rabies virus , recombinant dna , rabies , biology , virology , green fluorescent protein , recombinant virus , vector (molecular biology) , glycoprotein , complementary dna , virus , viral vector , rhabdoviridae , tracing , microbiology and biotechnology , gene , computer science , genetics , operating system

Recombinant rabies virus (RV) vectors expressing fluorescent proteins allow staining of neurons from many mammalian species and enable the study of neuron morphology. Because viral spread occurs only between neurons that have synaptic connections, these vectors also permit transsynaptic tracing. A recently established system for restriction of transsynaptic tracing to a single transsynaptic jump, dubbed monosynaptic tracing, uses glycoprotein gene-defective, pseudotyped RV. This allows infection of defined cells and transient complementation with the glycoprotein in situ to support a single step of transsynaptic crossing to presynaptic cells. Here, we introduce protocols describing the production of RV vectors, including the recovery of recombinant RV from complementary DNA (cDNA) and virus pseudotyping in vitro. This allows retrograde staining of neurons projecting to the inoculation site.

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