Skip to main content

Monosynaptic Tracing in Developing Circuits Using Modified Rabies Virus

  • Protocol
  • First Online:
Synapse Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1538))

Abstract

An attenuated rabies virus that expresses fluorescent protein has made it possible to analyze retrograde (presynaptic) monosynaptic connections in vivo. By combining attenuated rabies virus with a Cre-loxP based system to target cells in a subtype-specific fashion, it is possible to examine neuronal input in vivo onto any class of neuron, in development and in the mature brain. We describe here the methods to amplify deletion mutant, pseudotyped rabies virus, selectively target cells of interest using genetic and viral approaches, as well as the stereotaxic procedures required to target neuronal subtypes of interest in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 159.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Wickersham IR, Lyon DC, Barnard RJ, Mori T, Finke S, Conzelmann KK, Young JA, Callaway EM (2007) Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons. Neuron 53:639–647. doi:10.1016/j.neuron.2007.01.033, S0896-6273(07)00078-5 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Wall NR, Wickersham IR, Cetin A, De La Parra M, Callaway EM (2010) Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus. Proc Natl Acad Sci U S A 107:21848–21853. doi:10.1073/pnas.1011756107

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Larsen DD, Wickersham IR, Callaway EM (2007) Retrograde tracing with recombinant rabies virus reveals correlations between projection targets and dendritic architecture in layer 5 of mouse barrel cortex. Front Neural Circuits 1:5. doi:10.3389/neuro.04.005.2007

    PubMed  Google Scholar 

  4. Wickersham IR, Finke S, Conzelmann KK, Callaway EM (2007) Retrograde neuronal tracing with a deletion-mutant rabies virus. Nat Methods 4:47–49. doi:10.1038/nmeth999, nmeth999 [pii]

    Article  CAS  PubMed  Google Scholar 

  5. Sun Y, Nguyen AQ, Nguyen JP, Le L, Saur D, Choi J, Callaway EM, Xu X (2014) Cell-type-specific circuit connectivity of hippocampal CA1 revealed through cre-dependent rabies tracing. Cell Rep 7:269–280. doi:10.1016/j.celrep.2014.02.030

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Marshel JH, Mori T, Nielsen KJ, Callaway EM (2010) Targeting single neuronal networks for gene expression and cell labeling in vivo. Neuron 67:562–574. doi:10.1016/j.neuron.2010.08.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Kohara K, Pignatelli M, Rivest AJ, Jung H-Y, Kitamura T, Suh J, Frank D, Kajikawa K, Mise N, Obata Y, Wickersham IR, Tonegawa S (2014) Cell type-specific genetic and optogenetic tools reveal hippocampal CA2 circuits. Nat Neurosci 17:269–279. doi:10.1038/nn.3614

    Article  CAS  PubMed  Google Scholar 

  8. Lasek AW, Azouaou N (2010) Virus-delivered RNA interference in mouse brain to study addiction-related behaviors. Methods Mol Biol 602:283–298. doi:10.1007/978-1-60761-058-8_17

    Article  CAS  PubMed  Google Scholar 

  9. Weible AP, Schwarcz L, Wickersham IR, Deblander L, Wu H, Callaway EM, Seung HS, Kentros CG (2010) Transgenic targeting of recombinant rabies virus reveals monosynaptic connectivity of specific neurons. J Neurosci 30:16509–16513. doi:10.1523/JNEUROSCI.2442-10.2010 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Osakada F, Mori T, Cetin AH, Marshel JH, Virgen B, Callaway EM (2011) New rabies virus variants for monitoring and manipulating activity and gene expression in defined neural circuits. Neuron 71:617–631. doi:10.1016/j.neuron.2012.03.019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Srinivas S, Watanabe T, Lin CS, William CM, Tanabe Y, Jessell TM, Costantini F (2001) Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol 1:4. doi:10.1186/1471-213X-1-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Li Y, Stam FJ, Aimone JB, Goulding M, Callaway EM, Gage FH (2013) Molecular layer perforant path-associated cells contribute to feed-forward inhibition in the adult dentate gyrus. Proc Natl Acad Sci U S A 110:9106–9111. doi:10.1073/pnas.1306912110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Wang L, Sharma K, Deng H-X, Siddique T, Grisotti G, Liu E, Roos RP (2008) Restricted expression of mutant SOD1 in spinal motor neurons and interneurons induces motor neuron pathology. Neurobiol Dis 29:400–408. doi:10.1016/j.nbd.2007.10.004

    Article  CAS  PubMed  Google Scholar 

  14. Miyamichi K, Amat F, Moussavi F, Wang C, Wickersham I, Wall NR, Taniguchi H, Tasic B, Huang ZJ, He Z, Callaway EM, Horowitz MA, Luo L (2010) Cortical representations of olfactory input by trans-synaptic tracing. Nature 472:191–196. doi:10.1038/nature09714, nature09714 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Laura Cocas .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media LLC

About this protocol

Cite this protocol

Cocas, L., Fernandez, G. (2017). Monosynaptic Tracing in Developing Circuits Using Modified Rabies Virus. In: Poulopoulos, A. (eds) Synapse Development. Methods in Molecular Biology, vol 1538. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6688-2_23

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6688-2_23

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6686-8

  • Online ISBN: 978-1-4939-6688-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics