Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2015 Jun 17;35(24):8979-85.
doi: 10.1523/JNEUROSCI.0409-15.2015.

Monosynaptic Circuit Tracing with Glycoprotein-Deleted Rabies Viruses

Edward M Callaway  1 , Liqun Luo  2
Affiliations
Review

Monosynaptic Circuit Tracing with Glycoprotein-Deleted Rabies Viruses

Edward M Callaway et al. J Neurosci. .
No abstract available

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Glycoprotein deletion and EnvA pseudotyping of rabies virus. Normal rabies viral particles (top, left) are composed of a viral core surrounded by a host cell-derived membrane envelope in which the rabies glycoprotein (G; red spikes) is embedded. The core of the virus includes structural proteins and the negative strand RNA genome. The genome codes for five different proteins, including G (top, middle). The viral genome can be modified such that the coding sequence for G is deleted and can be replaced with the coding sequence for a transgene, such as GFP, which will be expressed in infected cells. G-deleted rabies virus particles (RVdG) are initially produced de novo from DNA and then can be propagated indefinitely (Osakada and Callaway, 2013). When propagated in cells that express G (data not shown), RVdG particles will bud out from the host cells with G on their envelope (G + RVdG, top, right). To pseudotype RVdG, cells expressing EnvA (blue lollipops) are infected with G + RVdG (bottom, left) and the particles that emerge will have EnvA on their envelope (bottom, right). These EnvA-pseudotyped particles are designated EnvA + RVdG.
Figure 2.
Figure 2.
Monosynaptic rabies tracing of inputs to Cre-expressing starter cells. Monosynaptic tracing with G-deleted rabies (RVdG) involves expression of the EnvA receptor, TVA, and rabies glycoprotein (G) in starter cells. These cells are subsequently infected specifically with EnvA + RVdG, which is trans-complemented by G to produce G + RVdG that spreads trans-synaptically to neurons providing synaptic input to starter cells. The spread of RVdG is monosynaptically restricted because the input cells lack G and G is required for trans-synaptic spread. In this example, the starter cells are selected on the basis of Cre expression in a mouse line expressing Cre in a cell type of interest. In the first step (1) a helper virus that expresses TVA, G, and a marker gene (RFP) in a Cre-dependent manner is injected in the location of desired starter cells. This results in expression of TVA, G, and RFP only in the Cre+ cells (green and red) (2). After a sufficient time for accumulation of TVA and G, EnvA + RVdG is injected in the location (3) and selectively infects the TVA+ starter cells. In this example the EnvA + RVdG contains the coding sequence for GFP resulting in GFP expression in the starter cells. The RVdG replicates in the starter cells and trans-complementation with G results in the production of G + RVdG that spreads trans-synaptically to input cells (4), which express GFP from the RV genome (green). The result is that starter cells are marked with both RFP and GFP while input cells express only GFP. Other neurons that do not express Cre and are not directly presynaptic to starter cells remain unlabeled (light yellow).
See this image and copyright information in PMC

References

    1. Arenkiel BR, Hasegawa H, Yi JJ, Larsen RS, Wallace ML, Philpot BD, Wang F, Ehlers MD. Activity-induced remodeling of olfactory bulb microcircuits revealed by monosynaptic tracing. PLoS One. 2011;6:e29423. doi: 10.1371/journal.pone.0029423. - DOI - PMC - PubMed
    1. Arnold DB. Actin and microtubule-based cytoskeletal cues direct polarized targeting of proteins in neurons. Sci Signal. 2009;2:pe49. doi: 10.1126/scisignal.283pe49. - DOI - PMC - PubMed
    1. Bauer A, Nolden T, Schröter J, Römer-Oberdörfer A, Gluska S, Perlson E, Finke S. Anterograde glycoprotein-dependent transport of newly generated rabies virus in dorsal root ganglion neurons. J Virol. 2014;88:14172–14183. doi: 10.1128/JVI.02254-14. - DOI - PMC - PubMed
    1. Bourane S, Grossmann KS, Britz O, Dalet A, Del Barrio MG, Stam FJ, Garcia-Campmany L, Koch S, Goulding M. Identification of a spinal circuit for light touch and fine motor control. Cell. 2015;160:503–515. doi: 10.1016/j.cell.2015.01.011. - DOI - PMC - PubMed
    1. Callaway EM. Transneuronal circuit tracing with neurotropic viruses. Curr Opin Neurobiol. 2008;18:617–623. doi: 10.1016/j.conb.2009.03.007. - DOI - PMC - PubMed

Publication types

LinkOut - more resources