Journal
NATURE PROTOCOLS
Volume 5, Issue 3, Pages 439-456Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.226
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Funding
- Deisseroth laboratory
- NIH NRSA
- SGF
- SIGF (Stanford Graduate Fellowships)
- Fonds National de la Recherche Scientifique
- NARSAD, NIH [K99]
- Fondation Leon Fredericq
- NIDA
- DARPA
- NARSAD
- William M. Keck Foundation
- Snyder Foundation
- Albert Yu and Mary Bechmann Foundation
- Wallace Coulter Foundation
- California Institute for Regenerative Medicine
- McKnight Foundation
- Esther A. and Joseph Klingenstein Fund
- NSF
- National Institute of Mental Health
- National Institute on Drug Abuse
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [F31NS059160, K99NS065009] Funding Source: NIH RePORTER
- OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH [DP1OD000616] Funding Source: NIH RePORTER
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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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