Three‐Photon Infrared Stimulation of Endogenous Neuroreceptors in Vivo

Author:

Sortino Rosalba12ORCID,Cunquero Marina3ORCID,Castro‐Olvera Gustavo3ORCID,Gelabert Ricard4ORCID,Moreno Miquel4ORCID,Riefolo Fabio125ORCID,Matera Carlo126ORCID,Fernàndez‐Castillo Noèlia2789ORCID,Agnetta Luca10,Decker Michael10,Lluch José M.411ORCID,Hernando Jordi4ORCID,Loza‐Alvarez Pablo3ORCID,Gorostiza Pau1212ORCID

Affiliation:

1. Institute for Bioengineering of Catalonia (IBEC) The Barcelona Institute for Science and Technology 08028 Barcelona Spain

2. CIBER-BBN ISCIII 28029 Madrid Spain

3. ICFO—Institut de Ciencies Fotoniques The Barcelona Institute of Science and Technology 08860 Castelldefels (Barcelona) Spain

4. Departament de Química Universitat Autònoma de Barcelona (UAB) 08193 Bellaterra Spain

5. Current address: Teamit Institute, Partnerships Barcelona Health Hub 08025 Barcelona Spain

6. Current address: Department of Pharmaceutical Sciences University of Milan 20133 Milan Italy

7. Departament de Genètica, Microbiologia i Estadística, Facultat de Biologia Universitat de Barcelona 08028 Barcelona Spain

8. Institut de Biomedicina de la Universitat de Barcelona (IBUB) 08028 Barcelona Spain

9. Institut de Recerca Sant Joan de Déu (IRSJD) 08950 Esplugues de Llobregat Spain

10. Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy and Food Chemistry Ludwig Maximilian University of Würzburg 97074 Würzburg Germany

11. Institut de Biotecnologia i de Biomedicina (IBB) UAB 08193 Bellaterra Spain

12. Catalan Institution of Research and Advanced Studies (ICREA) 08010 Barcelona Spain

Abstract

AbstractTo 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 and optogenetics. 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.

Funder

Agència de Gestió d'Ajuts Universitaris i de Recerca

Ministerio de Ciencia, Innovación y Universidades

'la Caixa' Foundation

European Social Fund

Ministerio de Economía y Competitividad

Publisher

Wiley

Subject

General Medicine

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