Transneuronal circuit tracing with neurotropic viruses

The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Current opinion in neurobiology (Impact Factor: 6.63). 05/2009; 18(6):617-23. DOI: 10.1016/j.conb.2009.03.007
Source: PubMed


Because neurotropic viruses naturally traverse neural pathways, they are extremely valuable for elucidating neural circuits. Naturally occurring herpes and rabies viruses have been used for transneuronal circuit tracing for decades. Depending on the type of virus and strain, virus can travel preferentially in the anterograde or the retrograde direction. More recently, genetic modifications have allowed for many improvements. These include: reduced pathogenicity; addition of marker genes; control of synaptic spread; pseudotyping for infection of selected cells; addition of ancillary genetic elements for combining circuit tracing with manipulation of activity or functional assays. These modifications, along with the likelihood of future developments, suggest that neurotropic viruses will be increasingly important and effective tools for future studies of neural circuits.

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    • "The dual-virus approach described here capitalizes upon the high efficiency and longterm tolerability of AAVs while also enabling robust optical control (Figures 2D and 2E) via catalytically driven activation of high copy-number transgenes. It is important to be aware that direct transduction of axon terminals with certain viruses can be achieved for targeting (reviewed in Callaway, 2008), and indeed virtually all viruses (including AAVs) carry some degree of this capability in a serotype-and circuit-dependent manner, as we and others have observed (Burger et al., 2004; Paterna et al., 2004; Callaway, 2008; Lima et al., 2009; Passini et al., 2005; Nathanson et al., 2009). It is also important to consider (in the process of selecting viruses) the need for high levels of infection and expression while avoiding adverse effects on transduced neurons over weeks to months for long-term electrophysiological or behavioral experiments. "
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    DESCRIPTION: Molecular and Cellular Approaches for Diversifying and Extending Optogenetics (Viviana Gradinaru,Feng Zhang) university of california , los anjeles and caltech
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    • "Lentivirus provides several advantages with spatially defined expression and combinations of tracers with optoge - netic and other functional constructs allowing examination of questions not previously possible using traditional tracing techniques . Analysis of neuronal circuitry has traditionally relied on neuronal tracers such as biotinylated dextran amine ( BDA ) , phaseolus vulgaris leucoagglutinin ( PHAL ) , wheat - germ agglutinin ( WGA ) , horseradish peroxidase ( HRP ) , cholera toxin , and more recently pseudorabies virus to label neuronal pathways ( Callaway , 2008 ; Huh et al . , 2010 ) . "
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    ABSTRACT: Lentiviruses have been extensively used as gene delivery vectors since the mid-1990s. Usually derived from the human immunodeficiency virus genome, they mediate efficient gene transfer to non-dividing cells, including neurons and glia in the adult mammalian brain. In addition, integration of the recombinant lentiviral construct into the host genome provides permanent expression, including the progeny of dividing neural precursors. In this review, we describe targeted vectors with modified envelope glycoproteins and expression of transgenes under the regulation of cell-selective and inducible promoters. This technology has broad utility to address fundamental questions in neuroscience and we outline how this has been used in rodents and primates. Combining viral tract tracing with immunohistochemistry and confocal or electron microscopy, lentiviral vectors provide a tool to selectively label and trace specific neuronal populations at gross or ultrastructural levels. Additionally, new generation optogenetic technologies can be readily utilized to analyze neuronal circuit and gene functions in the mature mammalian brain. Examples of these applications, limitations of current systems and prospects for future developments to enhance neuroscience knowledge will be reviewed. Finally, we will discuss how these vectors may be translated from gene therapy trials into the clinical setting.
    Frontiers in Molecular Neuroscience 04/2015; 8. DOI:10.3389/fnmol.2015.00014 · 4.08 Impact Factor
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    • "Although this property is known from the wild-type CVS strain of RABV (Callaway 2008), it has not been demonstrated for glycoprotein-deleted pseudotyped variants of the SAD B19 strain. To examine the ability of our anterograde RABV DG vector to confer the sparse labeling necessary for single neuron reconstruction, we injected 5–10 viral particles into the thalamic POm division of a 16-week-old mouse. "
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    ABSTRACT: Glycoprotein-deleted rabies virus (RABV ∆G) is a powerful tool for the analysis of neural circuits. Here, we demonstrate the utility of an anterograde RABV ∆G variant for novel neuroanatomical approaches involving either bulk or sparse neuronal populations. This technology exploits the unique features of RABV ∆G vectors, namely autonomous, rapid high-level expression of transgenes, and limited cytotoxicity. Our vector permits the unambiguous long-range and fine-scale tracing of the entire axonal arbor of individual neurons throughout the brain. Notably, this level of labeling can be achieved following infection with a single viral particle. The vector is effective over a range of ages (>14 months) aiding the studies of neurodegenerative disorders or aging, and infects numerous cell types in all brain regions tested. Lastly, it can also be readily combined with retrograde RABV ∆G variants. Together with other modern technologies, this tool provides new possibilities for the investigation of the anatomy and physiology of neural circuits.
    Brain Structure and Function 04/2014; 220(3). DOI:10.1007/s00429-014-0730-z · 5.62 Impact Factor
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