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Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures

Nature Protocols volume 5, pages 439–456 (2010)Cite this article

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Abstract

Elucidation of the neural substrates underlying complex animal behaviors depends on precise activity control tools, as well as compatible readout methods. Recent developments in optogenetics have addressed this need, opening up new possibilities for systems neuroscience. Interrogation of even deep neural circuits can be conducted by directly probing the necessity and sufficiency of defined circuit elements with millisecond-scale, cell type-specific optical perturbations, coupled with suitable readouts such as electrophysiology, optical circuit dynamics measures and freely moving behavior in mammals. Here we collect in detail our strategies for delivering microbial opsin genes to deep mammalian brain structures in vivo, along with protocols for integrating the resulting optical control with compatible readouts (electrophysiological, optical and behavioral). The procedures described here, from initial virus preparation to systems-level functional readout, can be completed within 4–5 weeks. Together, these methods may help in providing circuit-level insight into the dynamics underlying complex mammalian behaviors in health and disease.

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Figure 1: Optogenetic tools.
Figure 2: Stereotactic implantation of the cannula guide.
Figure 3: Preparation of the optical fiber for in vivo neural control in mammals.
Figure 4: Functional expression of microbial opsin genes in the rodent brain.

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Acknowledgements

We thank the entire Deisseroth laboratory for their support. F.Z. is supported by an NIH NRSA. V.G. is supported by SGF and SIGF (Stanford Graduate Fellowships). A.R.A. is supported by the Fonds National de la Recherche Scientifique, NARSAD, NIH (K99) and the Fondation Leon Fredericq. L.d.L. is supported by NIDA, DARPA and NARSAD. K.D. is supported by the William M. Keck Foundation, the Snyder Foundation, the Albert Yu and Mary Bechmann Foundation and the Wallace Coulter Foundation, as well as by California Institute for Regenerative Medicine, the McKnight Foundation, the Esther A. and Joseph Klingenstein Fund, NSF, National Institute of Mental Health, National Institute on Drug Abuse and the NIH Pioneer Award.

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Author notes

  1. Feng Zhang, Viviana Gradinaru and Antoine R Adamantidis: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Bioengineering, Stanford University, Stanford, California, USA

    Feng Zhang, Viviana Gradinaru, Remy Durand, Raag D Airan & Karl Deisseroth

  2. Program in Neuroscience, Stanford University, Stanford, California, USA

    Viviana Gradinaru

  3. Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, California, USA

    Antoine R Adamantidis, Luis de Lecea & Karl Deisseroth

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F.Z., V.G., A.R.A., R.D.A., R.D., L.d.L. and K.D. wrote the manuscript.

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Correspondence to Karl Deisseroth.

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Zhang, F., Gradinaru, V., Adamantidis, A. et al. Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures. Nat Protoc 5, 439–456 (2010). https://doi.org/10.1038/nprot.2009.226

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