To interrogate neural circuits and crack their codes, in vivo brain activity imaging must be combined with spatiotemporally precise stimulation in three dimensions using genetic or pharmacological specificity. This challenge requires deep penetration and focusing as provided by infrared light and multiphoton excitation, and has promoted two-photon photopharmacology[1] and optogenetics[2]. However, three- photon brain stimulation in vivo remains to be demonstrated. We report the regulation of neuronal activity in zebrafish larvae by three-photon excitation of a photoswitchable muscarinic agonist at 50 pM, a billion- fold lower concentration than used for uncaging, and with mid-infrared light of 1560 nm, the longest reported photoswitch wavelength. Robust, physiologically relevant photoresponses allow modulating brain activity in wild-type animals with spatiotemporal and pharmacological precision. Computational calculations predict that azobenzene-based ligands have high three-photon absorption cross-section and can be used directly with pulsed infrared light. The expansion of three-photon pharmacology will deeply impact basic neurobiology and neuromodulation phototherapies. [1] a)G. C. R. Ellis-Davies, Front Synaptic Neurosci 2018, 10, 48; b)F. Riefolo, C. Matera, A. Garrido-Charles, A. M. J. Gomila, R. Sortino, L. Agnetta, E. Claro, R. Masgrau, U. Holzgrabe, M. Batlle, M. Decker, E. Guasch, P. Gorostiza, J Am Chem Soc 2019, 141, 7628-7636; c)M. Izquierdo-Serra, M. Gascon-Moya, J. J. Hirtz, S. Pittolo, K. E. Poskanzer, E. Ferrer, R. Alibes, F. Busque, R. Yuste, J. Hernando, P. Gorostiza, J Am Chem Soc 2014, 136, 8693-8701; d)E. C. Carroll, S. Berlin, J. Levitz, M. A. Kienzler, Z. Yuan, D. Madsen, D. S. Larsen, E. Y. Isacoff, Proc Natl Acad Sci U S A 2015, 112, E776-785. [2] a)J. P. Rickgauer, D. W. Tank, Proc Natl Acad Sci U S A 2009, 106, 15025-15030; b)A. Begue, E. Papagiakoumou, B. Leshem, R. Conti, L. Enke, D. Oron, V. Emiliani, Biomed Opt Express 2013, 4, 2869- 2879; c)M. Klausen, M. Blanchard-Desce, Journal of Photochemistry and Photobiology C: Photochemistry Reviews 2021, 48, 100423.

Three-Photon Infrared Stimulation of Endogenous Neuroreceptors in Vivo / R. Sortino, M. Cunquero, G. Castro-Olvera, R. Gelabert, M. Moreno, F. Riefolo, C. Matera, N. Fernàndez-Castillo, L. Agnetta, M. Decker, J.M. Lluch, J. Hernando, P. Loza-Alvarez, P. Gorostiza. ((Intervento presentato al 29. convegno IUPAC Symposium on Photochemistry : 14 - 19 July tenutosi a Valencia nel 2024.

Three-Photon Infrared Stimulation of Endogenous Neuroreceptors in Vivo

F. Riefolo;C. Matera;
2024

Abstract

To interrogate neural circuits and crack their codes, in vivo brain activity imaging must be combined with spatiotemporally precise stimulation in three dimensions using genetic or pharmacological specificity. This challenge requires deep penetration and focusing as provided by infrared light and multiphoton excitation, and has promoted two-photon photopharmacology[1] and optogenetics[2]. However, three- photon brain stimulation in vivo remains to be demonstrated. We report the regulation of neuronal activity in zebrafish larvae by three-photon excitation of a photoswitchable muscarinic agonist at 50 pM, a billion- fold lower concentration than used for uncaging, and with mid-infrared light of 1560 nm, the longest reported photoswitch wavelength. Robust, physiologically relevant photoresponses allow modulating brain activity in wild-type animals with spatiotemporal and pharmacological precision. Computational calculations predict that azobenzene-based ligands have high three-photon absorption cross-section and can be used directly with pulsed infrared light. The expansion of three-photon pharmacology will deeply impact basic neurobiology and neuromodulation phototherapies. [1] a)G. C. R. Ellis-Davies, Front Synaptic Neurosci 2018, 10, 48; b)F. Riefolo, C. Matera, A. Garrido-Charles, A. M. J. Gomila, R. Sortino, L. Agnetta, E. Claro, R. Masgrau, U. Holzgrabe, M. Batlle, M. Decker, E. Guasch, P. Gorostiza, J Am Chem Soc 2019, 141, 7628-7636; c)M. Izquierdo-Serra, M. Gascon-Moya, J. J. Hirtz, S. Pittolo, K. E. Poskanzer, E. Ferrer, R. Alibes, F. Busque, R. Yuste, J. Hernando, P. Gorostiza, J Am Chem Soc 2014, 136, 8693-8701; d)E. C. Carroll, S. Berlin, J. Levitz, M. A. Kienzler, Z. Yuan, D. Madsen, D. S. Larsen, E. Y. Isacoff, Proc Natl Acad Sci U S A 2015, 112, E776-785. [2] a)J. P. Rickgauer, D. W. Tank, Proc Natl Acad Sci U S A 2009, 106, 15025-15030; b)A. Begue, E. Papagiakoumou, B. Leshem, R. Conti, L. Enke, D. Oron, V. Emiliani, Biomed Opt Express 2013, 4, 2869- 2879; c)M. Klausen, M. Blanchard-Desce, Journal of Photochemistry and Photobiology C: Photochemistry Reviews 2021, 48, 100423.
lug-2024
Settore CHIM/08 - Chimica Farmaceutica
Settore BIO/14 - Farmacologia
https://www.photoiupac2024.com/index.php
Three-Photon Infrared Stimulation of Endogenous Neuroreceptors in Vivo / R. Sortino, M. Cunquero, G. Castro-Olvera, R. Gelabert, M. Moreno, F. Riefolo, C. Matera, N. Fernàndez-Castillo, L. Agnetta, M. Decker, J.M. Lluch, J. Hernando, P. Loza-Alvarez, P. Gorostiza. ((Intervento presentato al 29. convegno IUPAC Symposium on Photochemistry : 14 - 19 July tenutosi a Valencia nel 2024.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/1094108
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