title: Anterograde or Retrograde Transsynaptic Circuit Tracing in Vertebrates with Vesicular Stomatitis Virus Vectors.
abstract: Viruses have been used as transsynaptic tracers, allowing one to map the inputs and outputs of neuronal populations, due to their ability to replicate in neurons and transmit in vivo only across synaptically connected cells. To date, their use has been largely restricted to mammals. In order to explore the use of such viruses in an expanded host range, we tested the transsynaptic tracing ability of recombinant vesicular stomatitis virus (rVSV) vectors in a variety of organisms. Successful infection and gene expression were achieved in a wide range of organisms, including vertebrate and invertebrate model organisms. Moreover, rVSV enabled transsynaptic tracing of neural circuitry in predictable directions dictated by the viral envelope glycoprotein (G), derived from either VSV or rabies virus (RABV). Anterograde and retrograde labeling, from initial infection and/or viral replication and transmission, was observed in Old and New World monkeys, seahorses, jellyfish, zebrafish, chickens, and mice. These vectors are widely applicable for gene delivery, afferent tract tracing, and/or directional connectivity mapping. Here, we detail the use of these vectors and provide protocols for propagating virus, changing the surface glycoprotein, and infecting multiple organisms using several injection strategies.
authors.affiliation: Department of Biology, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, California.|These authors contributed equally to this unit.
authors.forename: Kevin T
authors.initials: KT
authors.lastname: Beier
authors.affiliation: Department of Genetics, Department of Ophthalmology, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts.|These authors contributed equally to this unit.
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Mundell
authors.affiliation: Department of Neuroscience and Regenerative Medicine, Department of Neurology, James & Jean Culver Vision Discovery Institute, Medical College of Georgia, Augusta University, Augusta, Georgia.|These authors contributed equally to this unit.
authors.forename: Y Albert
authors.initials: YA
authors.lastname: Pan
authors.affiliation: Department of Genetics, Department of Ophthalmology, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Constance L
authors.initials: CL
authors.lastname: Cepko
mesh_terms: D000818:Animals; D002520:Cercopithecidae; D002645:Chickens; D003063:Cnidaria; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D008562:Membrane Glycoproteins; D051379:Mice; D056347:Neuroanatomical Tract-Tracing Techniques / Q000379:methods*; D051078:Platyrrhini; D021381:Protein Transport; D011820:Rabies virus / Q000502:physiology; D023701:Smegmamorpha; D013569:Synapses; D018116:Vesiculovirus / Q000235:genetics*; D014759:Viral Envelope Proteins; D015027:Zebrafish
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins
keywords: VSV; axon tracing; gene delivery; neural circuitry; transsynaptic tracing