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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References
Hikosaka, O. & Wurtz, R.H. Effects on eye movements of a GABA agonist and antagonist injected into monkey superior colliculus. Brain Res. 272, 368–372 (1983).
Boyden, E.S., Zhang, F., Bamberg, E., Nagel, G. & Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat. Neurosci. 8, 1263–1268 (2005).
Zhang, F., Wang, L.P., Boyden, E.S. & Deisseroth, K. Channelrhodopsin-2 and optical control of excitable cells. Nat. Methods 3, 785–792 (2006).
Zhang, F. et al. Multimodal fast optical interrogation of neural circuitry. Nature 446, 633–639 (2007).
Gradinaru, V., Thompson, K.R. & Deisseroth, K. eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications. Brain Cell Biol. 36, 129–139 (2008).
Gradinaru, V. et al. Targeting and readout strategies for fast optical neural control in vitro and in vivo. J. Neurosci. 27, 14231–14238 (2007).
Zhang, F. et al. Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures. Nat. Protoc. 5, 439–456 (2010).
Zhang, J. et al. Integrated device for optical stimulation and spatiotemporal electrical recording of neural activity in light-sensitized brain tissue. J. Neural Eng. 6, 055007 (2009).
Adamantidis, A.R., Zhang, F., Aravanis, A.M., Deisseroth, K. & de Lecea, L. Neural substrates of awakening probed with optogenetic control of hypocretin neurons. Nature 450, 420–424 (2007).
Aravanis, A.M. et al. An optical neural interface: in vivo control of rodent motor cortex with integrated fiberoptic and optogenetic technology. J. Neural Eng. 4, S143–S156 (2007).
Gradinaru, V., Mogri, M., Thompson, K.R., Henderson, J.M. & Deisseroth, K. Optical deconstruction of parkinsonian neural circuitry. Science 324, 354–359 (2009).
Airan, R.D., Thompson, K.R., Fenno, L.E., Bernstein, H. & Deisseroth, K. Temporally precise in vivo control of intracellular signaling. Nature 458, 1025–1029 (2009).
Tsai, H.C. et al. Phasic firing in dopaminergic neurons is sufficient for behavioral conditioning. Science 324, 1080–1084 (2009).
Han, X. et al. Millisecond-timescale optical control of neural dynamics in the nonhuman primate brain. Neuron 62, 191–198 (2009).
Gilja, V. et al. Challenges and opportunities for next-generation intra-cortically based neural prostheses. IEEE Trans. Biomed. Eng. (in the press).
Kim, S., Tathireddy, P., Normann, R.A. & Solzbacher, F. Thermal impact of an active 3-D microelectrode array implanted in the brain. IEEE Trans. Neural Syst. Rehabil. Eng. 15, 493–501 (2007).
Lee, J.H. et al. Global and local fMRI signals driven by neurons defined optogenetically by type and wiring. Nature 465, 788–792 (2010).
Cardin, J.A. et al. Targeted optogenetic stimulation and recording of neurons in vivo using cell type–specific expression of Channelrhodopsin-2. Nat. Protoc. 5, 247–254 (2010).
Gradinaru, V. et al. Molecular and cellular approaches for diversifying and extending optogenetics. Cell 141, 154–165 (2010).
Berndt, A., Yizhar, O., Gunaydin, L.A., Hegemann, P. & Deisseroth, K. Bi-stable neural state switches. Nat. Neurosci. 12, 229–234 (2009).
Bogen, I.L., Haug, K.H., Roberg, B., Fonnum, F. & Walaas, S.I. The importance of synapsin I and II for neurotransmitter levels and vesicular storage in cholinergic, glutamatergic and GABAergic nerve terminals. Neurochem. Int. 55, 13–21 (2009).
Caroni, P. Overexpression of growth-associated proteins in the neurons of adult transgenic mice. J. Neurosci. Methods 71, 3–9 (1997).
Adelsberger, H., Garaschuk, O. & Konnerth, A. Cortical calcium waves in resting newborn mice. Nat. Neurosci. 8, 988–990 (2005).
Disbrow, E., Litinas, E., Recanzone, G.H., Padberg, J. & Krubitzer, L. Cortical connections of the second somatosensory area and the parietal ventral area in macaque monkeys. J. Comp. Neurol. 462, 382–399 (2003).
Leichnetz, G.R. Afferent and efferent connections of the dorsolateral precentral gyrus (area 4, hand/arm region) in the macaque monkey, with comparisons to area 8. J. Comp. Neurol. 254, 460–492 (1986).
Burger, C. et al. Recombinant AAV viral vectors pseudotyped with viral capsids from serotypes 1, 2 and 5 display differential efficiency and cell tropism after delivery to different regions of the central nervous system. Mol. Ther. 10, 302–317 (2004).
Jones, E.G., Huntley, G.W. & Benson, D.L. Alpha calcium/calmodulin-dependent protein kinase II selectively expressed in a subpopulation of excitatory neurons in monkey sensory-motor cortex: comparison with GAD-67 expression. J. Neurosci. 14, 611–629 (1994).
Houser, C.R., Hendry, S.H., Jones, E.G. & Vaughn, J.E. Morphological diversity of immunocytochemically identified GABA neurons in the monkey sensory-motor cortex. J. Neurocytol. 12, 617–638 (1983).
Markram, H. et al. Interneurons of the neocortical inhibitory system. Nat. Rev. Neurosci. 5, 793–807 (2004).
Diester, I. & Nieder, A. Complementary contributions of prefrontal neuron classes in abstract numerical categorization. J. Neurosci. 28, 7737–7747 (2008).
Kaufman, M.T. et al. Roles of monkey premotor neuron classes in movement preparation and execution. J. Neurophysiol. 104, 799–810 (2010).
Calvin, W.H. & Sypert, G.W. Fast and slow pyramidal tract neurons: an intracellular analysis of their contrasting repetitive firing properties in the cat. J. Neurophysiol. 39, 420–434 (1976).
Mullen, R.J., Buck, C.R. & Smith, A.M. NeuN, a neuronal specific nuclear protein in vertebrates. Development 116, 201–211 (1992).
McLendon, R.E. & Bigner, D.D. Immunohistochemistry of the glial fibrillary acidic protein: basic and applied considerations. Brain Pathol. 4, 221–228 (1994).
Colle, M.A. et al. Efficient intracerebral delivery of AAV5 vector encoding human ARSA in non-human primate. Hum. Mol. Genet. 19, 147–158 (2010).
Dodiya, H.B. et al. Differential transduction following basal ganglia administration of distinct pseudotyped AAV capsid serotypes in nonhuman primates. Mol. Ther. 18, 579–587 (2010).
Kaplitt, M.G. et al. Safety and tolerability of gene therapy with an adeno-associated virus (AAV) borne GAD gene for Parkinson's disease: an open label, phase I trial. Lancet 369, 2097–2105 (2007).
Christine, C.W. et al. Safety and tolerability of putaminal AADC gene therapy for Parkinson disease. Neurology 73, 1662–1669 (2009).
Boutin, S. et al. Prevalence of serum IgG and neutralizing factors against adeno-associated virus types 1, 2, 5, 6, 8 and 9 in the healthy population: implications for gene therapy using AAV vectors. Hum. Gene Ther. 21, 704–712 (2010).
McFarland, N.R., Lee, J.S., Hyman, B.T. & McLean, P.J. Comparison of transduction efficiency of recombinant AAV serotypes 1, 2, 5, and 8 in the rat nigrostriatal system. J. Neurochem. 109, 838–845 (2009).
Frigault, M.M., Lacoste, J., Swift, J.L. & Brown, C.M. Live-cell microscopy—tips and tools. J. Cell Sci. 122, 753–767 (2009).
Nathanson, J.L., Yanagawa, Y., Obata, K. & Callaway, E.M. Preferential labeling of inhibitory and excitatory cortical neurons by endogenous tropism of adeno-associated virus and lentivirus vectors. Neuroscience 161, 441–450 (2009).
Grimm, D. et al. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J. Virol. 82, 5887–5911 (2008).
Gordon, T. et al. Accelerating axon growth to overcome limitations in functional recovery after peripheral nerve injury. Neurosurgery 65, A132–A144 (2009).
Chen, C.H. et al. Role of PKA in the anti-Thy-1 antibody-induced neurite outgrowth of dorsal root ganglionic neurons. J. Cell. Biochem. 101, 566–575 (2007).
Histed, M.H., Bonin, V. & Reid, R.C. Direct activation of sparse, distributed populations of cortical neurons by electrical microstimulation. Neuron 63, 508–522 (2009).
Gunaydin, L.A. et al. Ultrafast optogenetic control. Nat. Neurosci. 13, 387–392 (2010).
Taylor, D.M., Tillery, S.I. & Schwartz, A.B. Direct cortical control of 3D neuroprosthetic devices. Science 296, 1829–1832 (2002).
Churchland, M.M. & Shenoy, K.V. Delay of movement caused by disruption of cortical preparatory activity. J. Neurophysiol. 97, 348–359 (2007).
Noudoost, B. & Moore, T. A reliable microinjectrode system for use in behaving monkeys. J. Neurosci. Methods 194, 218–223 (2011).
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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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.
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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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DOI: https://doi.org/10.1038/nn.2749
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