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An optogenetic toolbox designed for primates

Nature Neuroscience volume 14, pages 387–397 (2011)Cite this article

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Abstract

Optogenetics is a technique for controlling subpopulations of neurons in the intact brain using light. This technique has the potential to enhance basic systems neuroscience research and to inform the mechanisms and treatment of brain injury and disease. Before launching large-scale primate studies, the method needs to be further characterized and adapted for use in the primate brain. We assessed the safety and efficiency of two viral vector systems (lentivirus and adeno-associated virus), two human promoters (human synapsin (hSyn) and human thymocyte-1 (hThy-1)) and three excitatory and inhibitory mammalian codon-optimized opsins (channelrhodopsin-2, enhanced Natronomonas pharaonis halorhodopsin and the step-function opsin), which we characterized electrophysiologically, histologically and behaviorally in rhesus monkeys (Macaca mulatta). We also introduced a new device for measuring in vivo fluorescence over time, allowing minimally invasive assessment of construct expression in the intact brain. We present a set of optogenetic tools designed for optogenetic experiments in the non-human primate brain.

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Figure 1: Schematic overview of preparation.
Figure 2: Representative example of electrophysiology results and population summary from the eNpHR2.0-expressing site.
Figure 3: Representative examples of electrophysiology results and population summary from the ChR2-expressing sites.
Figure 4: Lack of effect of optical stimulation on passive behavior.
Figure 5: Prolonged activation of spiking with a step function opsin (SFO).
Figure 6: In vivo fluorescence detector and measurements.
Figure 7: Histological analysis of cortex.
Figure 8: Evaluation of cell health.

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Acknowledgements

We thank M. Vessal and A. Lilak for technical assistance during perfusions, M. Churchland for technical support during surgeries and technical discussions, S. Ryu for surgical assistance, M. Risch for veterinary care, D. Haven for technical support, S. Eisensee for administrative assistance, I. Witten for advice and members of the Deisseroth and Shenoy laboratories for discussions. I.D. is supported by a Human Frontier Science Program fellowship and a German Academic Exchange Service Award, M.T.K. by a National Science Foundation graduate fellowship, M.M. by Bio-X and a Stanford Graduate Fellowship and R.P. by a Stanford University Dean's Postdoctoral Fellowship Award. W.G. is supported by a Stanford Graduate Fellowship. O.Y. is supported by a Human Frontier Science Program fellowship. K.D. is supported by the William M. Keck Foundation, the Snyder Foundation, the Albert Yu and Mary Bechmann Foundation, the Wallace Coulter Foundation, the California Institute for Regenerative Medicine, the McKnight Foundation, the Esther A. and Joseph Klingenstein Fund, the National Science Foundation, the National Institute of Mental Health, the National Institute on Drug Abuse and a US National Institutes of Health Director's Pioneer Award. K.V.S. is supported by a Burroughs Wellcome Fund Career Award in the Biomedical Sciences, US National Institutes of Health–National Institute of Neurological Disorders and Stroke grant CRCNS R01-NS054283 and a US National Institutes of Health Director's Pioneer Award (1DP1OD006409). K.D. and K.V.S. are supported by Defense Advanced Research Projects Agency Reorganization and Plasticity to Accelerate Injury Recovery (N66001-10-C-2010).

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Author notes

  1. Ramin Pashaie

    Present address: Present address: Department of Electrical Engineering and Computer Science, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin, USA.,

Authors and Affiliations

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

    Ilka Diester, Murtaza Mogri, Ramin Pashaie, Werapong Goo, Ofer Yizhar, Charu Ramakrishnan, Karl Deisseroth & Krishna V Shenoy

  2. Neurosciences Program, Stanford University, Stanford, California, USA

    Matthew T Kaufman, Karl Deisseroth & Krishna V Shenoy

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

    Karl Deisseroth

  4. Howard Hughes Medical Institute, Stanford University, Stanford, California, USA

    Karl Deisseroth

  5. Department of Electrical Engineering, Stanford University, Stanford, California, USA

    Krishna V Shenoy

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Contributions

I.D., K.V.S. and K.D. conceived and designed the experiments. I.D. wrote the manuscript and all authors contributed to its editing. I.D. conducted all experiments, histological analysis and data analysis. M.T.K. contributed to the neural recording and stimulation experiments and their analysis. R.P. developed the in vivo fluorescence detector and M.M. contributed to the in vivo fluorescence measurements and analysis. W.G. participated in immunostaining. C.R. designed and cloned the hThy-1 and hSyn constructs. O.Y. provided the SFO viral vector. K.D. and K.V.S. supervised all aspects of the work.

Corresponding authors

Correspondence to Karl Deisseroth or Krishna V Shenoy.

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Diester, I., Kaufman, M., Mogri, M. et al. An optogenetic toolbox designed for primates. Nat Neurosci 14, 387–397 (2011). https://doi.org/10.1038/nn.2749

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