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Retrograde neuronal tracing with a deletion-mutant rabies virus

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Abstract

We have constructed a deletion-mutant rabies virus encoding EGFP and find it to be an excellent tool for studying detailed morphology and physiology of neurons projecting to injection sites within the mammalian brain. The virus cannot spread beyond initially infected cells yet, unlike other viral vectors, replicates its core within them. The cells therefore fluoresce intensely, revealing fine dendritic and axonal structure with no background from partially or faintly labeled cells.

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Figure 1: The recombinant rabies virus SADΔG-EGFP.
Figure 2: Comparison of SADΔG-EGFP and lentivirus.

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Acknowledgements

We thank M. De La Parra and K. Roby for technical assistance, G. Tiscornia and I. Verma (Salk Institute) for plasmids and assistance with making the lentivirus, and D. Chambers for assistance with confocal microscopy and fluorescence quantification. We also thank L. Enquist (Princeton Univ.) for stocks of pseudorabies virus and S. Young (Salk Institute) for providing adenovirus. The SAD B19 glycoprotein expression plasmid pHCMV-RabiesG was a gift from M. Sena-Esteves (Harvard Medical School). This work was supported by US National Institute of Mental Health grant no. 63912 and Deutsche Forschungsgemeinschaft grant no. SFB455-A3.

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Author notes

  1. Stefan Finke

    Present address: Friedrich Loeffler Institute, Federal Research Institute for Animal Health, Boddenblick 5a, 17493, Greifswald-Insel Riems, Germany

Authors and Affiliations

  1. Systems Neurobiology Laboratory, The Salk Institute for Biological Studies, 10010 North Torrey Pines Rd., La Jolla, 92037, California, USA

    Ian R Wickersham & Edward M Callaway

  2. Max von Pettenkofer Institute & Gene Center, Ludwig Maximilians University, Feodor-Lynen-Str. 25, Muenchen, D-81377, Germany

    Stefan Finke & Karl-Klaus Conzelmann

Authors
  1. Ian R Wickersham
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  2. Stefan Finke
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  3. Karl-Klaus Conzelmann
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  4. Edward M Callaway
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Corresponding author

Correspondence to Ian R Wickersham.

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Competing interests

Generation of non-segmented negative-stranded RNA viruses from cloned DNA, described in this paper, is covered by US patent number 6,033,886, “Recombinant infectious non-segmented negative strand RNA virus,” filed Feb. 28, 1997; inventor: K.K.C.

Supplementary information

Supplementary Fig. 1

High EGFP expression reveals fine structural details of infected cells without immunohistochemical amplification.

Supplementary Fig. 2

Survival time course of neurons infected with SADΔG-EGFP.

Supplementary Methods

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Wickersham, I., Finke, S., Conzelmann, KK. et al. Retrograde neuronal tracing with a deletion-mutant rabies virus. Nat Methods 4, 47–49 (2007). https://doi.org/10.1038/nmeth999

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