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Virally mediated optogenetic excitation and inhibition of pain in freely moving nontransgenic mice

Abstract

Primary nociceptors are the first neurons involved in the complex processing system that regulates normal and pathological pain1. Because of constraints on pharmacological and electrical stimulation, noninvasive excitation and inhibition of these neurons in freely moving nontransgenic animals has not been possible. Here we use an optogenetic2 strategy to bidirectionally control nociceptors of nontransgenic mice. Intrasciatic nerve injection of adeno-associated viruses encoding an excitatory opsin enabled light-inducible stimulation of acute pain, place aversion and optogenetically mediated reductions in withdrawal thresholds to mechanical and thermal stimuli. In contrast, viral delivery of an inhibitory opsin enabled light-inducible inhibition of acute pain perception, and reversed mechanical allodynia and thermal hyperalgesia in a model of neuropathic pain. Light was delivered transdermally, allowing these behaviors to be induced in freely moving animals. This approach may have utility in basic and translational pain research, and enable rapid drug screening and testing of newly engineered opsins.

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Figure 1: Intrasciatic injection of AAV6-hSyn-ChR2-eYFP results in transduction of unmyelinated nociceptors projecting to spinal cord lamina I/IIo.
Figure 2: Transdermal illumination of AAV6-hSyn-ChR2-eYFP–injected mice results in tunable pain-like behavior and sensitizes mice to mechanical and thermal stimuli.
Figure 3: Transdermal illumination of AAV6-hSyn-NpHR-eYFP–injected mice desensitizes mice to mechanical and thermal stimuli and reverses mechanical allodynia and thermal hyperalgesia caused by a chronic constriction injury (CCI).

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Acknowledgements

We thank H. Liske, X. Qian, S. Mackey, A. Weitz, D.J. Clark, D.C. Yeomans, P. Sabhaie, C. Gorini, S. Young and H. Scutt for useful discussions and assistance with experiments. This study was supported by the US National Institutes of Health (National Institute of Neurological Disorders and Stroke grant R01-NS080954), the Stanford Bio-X NeuroVentures program and the Stanford Bio-X Interdisciplinary Initiatives program. S.M.I. was supported by an Office of Technology Licensing Stanford Graduate Fellowship and by a Howard Hughes Medical Institute International Student Research Fellowship. K.L.M. was supported by a Bio-X Bioengineering Graduate Fellowship and by a Stanford Interdisciplinary Graduate Fellowship. C.T. was supported by a Swiss National Science Foundation Fellowship.

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S.M.I., K.L.M., C.T. and S.L.D. designed the experiments. S.M.I. and K.L.M. performed the experiments. C.R. performed cell culture and created vectors. S.Y.L. performed electrophysiology. K.D. contributed reagents and tools. S.M.I., K.L.M. and S.L.D. wrote and edited the paper, with comments from all other authors.

Corresponding author

Correspondence to Scott L Delp.

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Competing interests

C.T., K.D., and S.L.D. have a financial interest in Circuit Therapeutics, Inc., which, however, did not support this work. S.M.I., K.L.M., C.T., K.D. and S.L.D. have disclosed these findings to the Stanford Office of Technology Licensing for potential use in the identification of new treatments for pain.

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Supplementary Figures 1–8 (PDF 4611 kb)

Supplementary Video 1

Video demonstration of transdermal optogenetic activation of nociceptors. A mouse with bilateral ChR2 expression is placed in a clear cylinder on a transparent glass plate, and allowed to habituate to its environment. When blue light (473 nm, 1 mW/mm2) is shone on the plantar surface of the mouse's skin it immediately withdraws its paw, engages in prolonged licking behavior, and shakes its paw. However, when yellow light (593 nm, 1 mW/mm2) is shone on the skin, there is no observable change in behavior, and the mouse moves around normally. Also, when blue light (473 nm, 1 mW/mm2) is shone on the paw of a bilaterally injected YFP+ mouse, no change in behavior is seen, and the mouse moves around normally. (MOV 8136 kb)

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Iyer, S., Montgomery, K., Towne, C. et al. Virally mediated optogenetic excitation and inhibition of pain in freely moving nontransgenic mice. Nat Biotechnol 32, 274–278 (2014). https://doi.org/10.1038/nbt.2834

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