Protocol | Published:

Targeted optogenetic stimulation and recording of neurons in vivo using cell-type-specific expression of Channelrhodopsin-2

Nature Protocols volume 5, pages 247254 (2010) | Download Citation

Abstract

A major long-term goal of systems neuroscience is to identify the different roles of neural subtypes in brain circuit function. The ability to causally manipulate selective cell types is critical to meeting this goal. This protocol describes techniques for optically stimulating specific populations of excitatory neurons and inhibitory interneurons in vivo in combination with electrophysiology. Cell type selectivity is obtained using Cre-dependent expression of the light-activated channel Channelrhodopsin-2. We also describe approaches for minimizing optical interference with simultaneous extracellular and intracellular recording. These optogenetic techniques provide a spatially and temporally precise means of studying neural activity in the intact brain and allow a detailed examination of the effect of evoked activity on the surrounding local neural network. Injection of viral vectors requires 30–45 min, and in vivo electrophysiology with optogenetic stimulation requires 1–4 h.

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Acknowledgements

We thank members of the Tsai and Moore laboratories for discussions and comments on the paper and M.J. Higley for help with optics. This study was supported by grants to C.I.M. from Tom F. Petersen, the NIH and the NSF, and by the Simons Foundation Autism Research Initiative to L.-H.T. K.D. was supported by the NIH Pioneer Program. L.-H.T. is an Investigator of the Howard Hughes Medical Institute. J.A.C. was supported by a K99 from the NIH/NEI; M.C. and K.M. by postdoctoral fellowships from the Knut och Alice Wallenberg Foundation; M.C. by a NARSAD Young Investigator Award; and F.Z. by an NIH NRSA.

Author information

Author notes

    • Jessica A Cardin
    •  & Marie Carlén

    These authors contributed equally to this work.

Affiliations

  1. McGovern Institute for Brain Research and Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA.

    • Jessica A Cardin
    • , Ulf Knoblich
    •  & Christopher I Moore
  2. Department of Neuroscience, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

    • Jessica A Cardin
  3. Picower Institute for Learning and Memory, Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA.

    • Marie Carlén
    • , Konstantinos Meletis
    •  & Li-Huei Tsai
  4. Stanley Center for Psychiatric Research, Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Marie Carlén
    • , Konstantinos Meletis
    •  & Li-Huei Tsai
  5. Department of Bioengineering, Stanford University, Stanford, California, USA.

    • Feng Zhang
    •  & Karl Deisseroth
  6. Howard Hughes Medical Institute, Cambridge, Massachusetts, USA.

    • Li-Huei Tsai

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Contributions

F.Z. and K.D. designed and cloned the AAV DIO ChR2-mCherry vector; M.C. and K.M. characterized the virus in vitro and in vivo and injected the animals; M.C. performed histological analyses; J.A.C. developed the experimental paradigm, and performed and analyzed the extracellular recordings; U.K. and J.A.C. performed the intracellular recordings; U.K. analyzed the intracellular data; and J.A.C., M.C., K.M., U.K., L.-H.T., and C.I.M. wrote the paper.

Corresponding authors

Correspondence to Karl Deisseroth or Li-Huei Tsai or Christopher I Moore.

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DOI

https://doi.org/10.1038/nprot.2009.228

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