Retrograde neuronal tracing with a deletion-mutant rabies virus (original) (raw)
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- Published: 10 December 2006
Nature Methods volume 4, pages 47–49 (2007)Cite this article
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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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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
- Stefan Finke
Present address: Friedrich Loeffler Institute, Federal Research Institute for Animal Health, Boddenblick 5a, 17493, Greifswald-Insel Riems, Germany
Authors and Affiliations
- 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 - Max von Pettenkofer Institute & Gene Center, Ludwig Maximilians University, Feodor-Lynen-Str. 25, Muenchen, D-81377, Germany
Stefan Finke & Karl-Klaus Conzelmann
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- Ian R Wickersham
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Correspondence toIan 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.
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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
- Received: 30 May 2006
- Accepted: 30 October 2006
- Published: 10 December 2006
- Issue Date: 01 January 2007
- DOI: https://doi.org/10.1038/nmeth999