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Published February 18, 2010 | Accepted Version
Journal Article Open

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

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.

Additional Information

© 2010 Nature Publishing Group. Published online: 18 February 2010. 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. Author Contributions: 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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