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Surgical implantation of wireless, battery-free optoelectronic epidural implants for optogenetic manipulation of spinal cord circuits in mice

Nature Protocols volume 16, pages 3072–3088 (2021)Cite this article

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Abstract

The use of optogenetics to regulate neuronal activity has revolutionized the study of the neural circuitry underlying a number of complex behaviors in rodents. Advances have been particularly evident in the study of brain circuitry and related behaviors, while advances in the study of spinal circuitry have been less striking because of technical hurdles. We have developed and characterized a wireless and fully implantable optoelectronic device that enables optical manipulation of spinal cord circuitry in mice via a microscale light-emitting diode (µLED) placed in the epidural space (NeuroLux spinal optogenetic device). This protocol describes how to surgically implant the device into the epidural space and then analyze light-induced behavior upon µLED activation. We detail optimized optical parameters for in vivo stimulation and demonstrate typical behavioral effects of optogenetic activation of nociceptive spinal afferents using this device. This fully wireless spinal µLED system provides considerable versatility for behavioral assays compared with optogenetic approaches that require tethering of animals, and superior temporal and spatial resolution when compared with other methods used for circuit manipulation such as chemogenetics. The detailed surgical approach and improved functionality of these spinal optoelectronic devices substantially expand the utility of this approach for the study of spinal circuitry and behaviors related to mechanical and thermal sensation, pruriception and nociception. The surgical implantation procedure takes ~1 h. The time required for the study of behaviors that are modulated by the light-activated circuit is variable and will depend upon the nature of the study.

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Fig. 1: Schematics of the updated spinal optogenetic device.
Fig. 2: Graphical representation of the surgical procedure for implantation of the spinal µLED device.
Fig. 3: Electrophysiological characterization of light-induced activation of TrpV1+ primary afferents in an ex vivo spinal cord slice preparation.
Fig. 4: Procedure for implantation of the spinal µLED device.
Fig. 5: General effects of spinal optogenetic device implantation on mice.
Fig. 6: Light-evoked pain-like behavior in TRPV1-ChR2 mice.
Fig. 7: Effects of opioid administration on light-evoked pain-like behavior.

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Source data are provided with this paper. Additional requests should be addressed to the corresponding authors.

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Acknowledgements

We thank J. Sinn-Hanlon for generating the illustrations in Fig. 2. This work was supported by NIH grants NS042595 to R.G., NS103472-02 to J.G.R., DA049569 to B.C. and NEUROLUX SBIR GRANT 5R44MH114944-02.

Author information

Authors and Affiliations

  1. Washington University Pain Center and Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO, USA

    Jose G. Grajales-Reyes, Bryan A. Copits, Sherri K. Vogt, Robert W. Gereau IV & Judith P. Golden

  2. Medical Scientist Training Program, Washington University in St. Louis, St. Louis, MO, USA

    Jose G. Grajales-Reyes

  3. NeuroLux Inc., Evanston, IL, USA

    Ferrona Lie, Yongjoon Yu & Anthony R. Banks

  4. Department of Mechanical Engineering, Evanston, IL, USA

    Ferrona Lie, Raudel Avila & Yonggang Huang

  5. Department of Materials Science and Engineering, Northwestern University, Evanston, IL, USA

    Yonggang Huang, Anthony R. Banks & John A. Rogers

  6. Department of Neuroscience and Affiliated Faculty, Northwestern University, Evanston, IL, USA

    Yonggang Huang, Anthony R. Banks & John A. Rogers

Authors
  1. Jose G. Grajales-Reyes
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Contributions

J.P.G. and R.W.G. developed the surgical approach. B.A.C. performed electrophysiology experiments. J.G.G. performed sensory behavior experiments. J.G.G. and J.P.G. performed optogenetic experiments. S.K.V. maintained and genotyped mice used in these studies. R.A. and Y.H. provided support with computer modeling for µLED device functioning. J.A.R, A.R.B., F.L. and Y.Y. designed and implemented all of the updates to the spinal µLED device in addition to providing technical support. All authors contributed to writing and editing the manuscript.

Corresponding authors

Correspondence to Robert W. Gereau IV or Judith P. Golden.

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Competing interests

A.R.B., R.W.G. and J.A.R. are cofounders of NeuroLux. F.L. and Y.Y. work for NeuroLux, a company that manufactures wireless optoelectronic devices. The device described here is included in the current NeuroLux portfolio29.

Additional information

Peer review information Nature Protocols thanks Maysam Ghovanloo, Ryan Koppes and the other, anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Related links

Key references using this protocol

Samineni, V. K. et al. Pain 158, 2108–2116 (2017): https://doi.org/10.1097/j.pain.0000000000000968

Park, S. I. et al. Nat. Biotechnol. 33, 1280–1286 (2015): https://doi.org/10.1038/nbt.3415

Vázquez-Guardado, A. et al. Nat. Neurosci. 23, 1522–1536 (2020): https://doi.org/10.1038/s41593-020-00739-8

Zhang, Y. et al Sci. Adv. 5, eaaw5296 (2019): https://doi.org/10.1126/sciadv.aaw5296

Supplementary information

Supplementary Information

Supplementary Methods and Supplementary Figs. 1–6.

Reporting Summary

Supplementary Video 1

Step-by-step demonstration of the surgical implantation procedure for the spinal µLED device.

Source data

Source Data Fig. 3

Statistical source data for physiology and animal behavior.

Source Data Fig. 5

Statistical source data for physiology and animal behavior.

Source Data Fig. 6

Statistical source data for physiology and animal behavior.

Source Data Fig. 7

Statistical source data for physiology and animal behavior.

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Grajales-Reyes, J.G., Copits, B.A., Lie, F. et al. Surgical implantation of wireless, battery-free optoelectronic epidural implants for optogenetic manipulation of spinal cord circuits in mice. Nat Protoc 16, 3072–3088 (2021). https://doi.org/10.1038/s41596-021-00532-2

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