Viral strategies for studying the brain, including a replication-restricted self-amplifying delta-G vesicular stomatis virus that rapidly expresses transgenes in brain and can generate a multicolor golgi-like expression.

Full Abstract

Viruses have substantial value as vehicles for transporting transgenes into neurons. Each virus has its own set of attributes for addressing neuroscience-related questions. Here we review some of the advantages and limitations of herpes, pseudorabies, rabies, adeno-associated, lentivirus, and others to study the brain. We then explore a novel recombinant vesicular stomatitis virus (dG-VSV) with the G-gene deleted and transgenes engineered into the first position of the RNA genome, which replicates only in the first brain cell infected, as corroborated with ultrastructural analysis, eliminating spread of virus. Because of its ability to replicate rapidly and to express multiple mRNA copies and additional templates for more copies, reporter gene expression is amplified substantially, over 500-fold in 6 hours, allowing detailed imaging of dendrites, dendritic spines, axons, and axon terminal fields within a few hours to a few days after inoculation. Green fluorescent protein (GFP) expression is first detected within 1 hour of inoculation. The virus generates a Golgi-like appearance in all neurons or glia of regions of the brain tested. Whole-cell patch-clamp electrophysiology, calcium digital imaging with fura-2, and time-lapse digital imaging showed that neurons appeared physiologically normal after expressing viral transgenes. The virus has a wide range of species applicability, including mouse, rat, hamster, human, and Drosophila cells. By using dG-VSV, we show efferent projections from the suprachiasmatic nucleus terminating in the periventricular region immediately dorsal to the nucleus. DG-VSVs with genes coding for different color reporters allow multicolor visualization of neurons wherever applied.

Author information

Author/s: van den Pol, Anthony N (AN); Ozduman, Koray (K); Wollmann, Guido (G); Ho, Winson S C (WS); Simon, Ian (I); Yao, Yang (Y); Rose, John K (JK); Ghosh, Prabhat (P);

Affiliation: Department of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut 06520, USA. anthony.vandenpol(-atsign-)yale.edu

Grants: A1/NS48854 (Agency:NINDS NIH HHS) ; CA124737 (Agency:NCI NIH HHS) ; NS34887 (Agency:NINDS NIH HHS) ; NS41454 (Agency:NINDS NIH HHS) ; NS48476 (Agency:NINDS NIH HHS)

Journal and publication information

Publication Type: Journal Article; Research Support, N.I.H., Extramural

Journal: The Journal of comparative neurology (J Comp Neurol), published in United States. (Language: eng)

Reference: 2009-Oct; vol 516 (issue 6) : pp 456-81

Dates: Created 2009/08/20; Completed 2009/10/26;

PMID: 19672982, status: MEDLINE (last retrieval date: 10/26/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Animals
Brain - physiology; ultrastructure; virology
Cells, Cultured
Gene Transfer Techniques
Genes, Reporter
Genetic Vectors
Humans
Membrane Glycoproteins - genetics
Mice

Neuroglia - ultrastructure; virology
Neurons - physiology; ultrastructure; virology
RNA, Messenger - metabolism
RNA, Viral - genetics
Transgenes
Vesiculovirus - genetics; physiology
Viral Envelope Proteins - genetics
Virus Replication
Viruses - genetics

Associated Chemicals: G protein, vesicular stomatitis virus (0) ; Membrane Glycoproteins (0) ; RNA, Messenger (0) ; RNA, Viral (0) ; Viral Envelope Proteins (0)

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