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In situ electrochemical generation of nitric oxide for neuronal modulation

Abstract

Understanding the function of nitric oxide, a lipophilic messenger in physiological processes across nervous, cardiovascular and immune systems, is currently impeded by the dearth of tools to deliver this gaseous molecule in situ to specific cells. To address this need, we have developed iron sulfide nanoclusters that catalyse nitric oxide generation from benign sodium nitrite in the presence of modest electric fields. Locally generated nitric oxide activates the nitric oxide-sensitive cation channel, transient receptor potential vanilloid family member 1 (TRPV1), and the latency of TRPV1-mediated Ca2+ responses can be controlled by varying the applied voltage. Integrating these electrocatalytic nanoclusters with multimaterial fibres allows nitric oxide-mediated neuronal interrogation in vivo. The in situ generation of nitric oxide in the ventral tegmental area with the electrocatalytic fibres evoked neuronal excitation in the targeted brain region and its excitatory projections. This nitric oxide generation platform may advance mechanistic studies of the role of nitric oxide in the nervous system and other organs.

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Fig. 1: Fe3S4 and Pt-Fe3S4 nanocatalysts for the electrochemical reduction of NO2 to NO.
Fig. 2: Electrochemically produced NO triggers TRPV1.
Fig. 3: Signalling pathways mediated by electrocatalytic NO generation in vitro.
Fig. 4: Fabrication and characterization of the NO delivery fibre.
Fig. 5: Neuronal stimulation mediated by NO delivery through implanted fibres in vivo.

Data availability

The data that support the plots within this paper and other findings of this study are available from the corresponding authors upon reasonable request.

Code availability

All scripts are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank D. Kim and F. Zhang for the generous gifts of the plasmids and cell lines. This work was funded in part by the National Institute of Neurological Disorders and Stroke (5R01NS086804) and the National Institutes of Health (NIH) BRAIN Initiative (1R01MH111872). This work made use of the MIT MRSEC Shared Experimental Facilities under award number DMR-14-19807 from the National Science Foundation (NSF). Funding for this research was also provided by the Department of Chemical Engineering at MIT. J.P. is a recipient of a scholarship from the Kwanjeong Educational Foundation. J.H.M. and Z.J.S. are supported by NSF Graduate Research Fellowships under grant number 1122374. T.T. was supported by the NEC Corporation. S.R. acknowledges funding support from the NIH Pathway to Independence Award (National Institute of Mental Health 1K99MH120279-01) and a grant from the Simons Foundation to the Simons Center for the Social Brain at MIT.

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J.P., K.J., K.M. and P.A. designed all experiments and performed all analyses. K.J. and J.H.M. synthesized the nanocatalysts and evaluated their electrocatalytic activity for NO generation. K.J. and Z.J.S. calculated diffusion profiles of NO. A.S., T.T., and T.K. fabricated the NO-delivery fibre. Y.F. offered insights into fibre fabrication. J.P., K.J. and P.C. performed in vitro TRPV1 modulation using the electrochemical NO delivery system. J.P. and P.C. conducted in vivo neuronal stimulation with the NO-delivery fibre. J.P., S.R. and D.R. performed immunohistochemistry analyses. J.P., F.K. and O.Y. conducted the electrophysiology analyses. J.P. and D.R. performed the cGMP assays. All co-authors contributed to the writing of the manuscript.

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Correspondence to Karthish Manthiram or Polina Anikeeva.

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Supplementary methods, Figs. 1–29 and refs. 1–7.

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Park, J., Jin, K., Sahasrabudhe, A. et al. In situ electrochemical generation of nitric oxide for neuronal modulation. Nat. Nanotechnol. 15, 690–697 (2020). https://doi.org/10.1038/s41565-020-0701-x

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