Rationally designed azobenzene photoswitches for efficient two-photon neuronal excitation.
Gisela CabréAida Garrido-CharlesMiquel MorenoMiquel BoschMontserrat Porta-de-la-RivaMichael KriegMarta Gascón-MoyaNúria CamareroRicard GelabertJosé M LluchFélix BusquéJordi HernandoPau GorostizaRamon AlibésPublished in: Nature communications (2019)
Manipulation of neuronal activity using two-photon excitation of azobenzene photoswitches with near-infrared light has been recently demonstrated, but their practical use in neuronal tissue to photostimulate individual neurons with three-dimensional precision has been hampered by firstly, the low efficacy and reliability of NIR-induced azobenzene photoisomerization compared to one-photon excitation, and secondly, the short cis state lifetime of the two-photon responsive azo switches. Here we report the rational design based on theoretical calculations and the synthesis of azobenzene photoswitches endowed with both high two-photon absorption cross section and slow thermal back-isomerization. These compounds provide optimized and sustained two-photon neuronal stimulation both in light-scattering brain tissue and in Caenorhabditis elegans nematodes, displaying photoresponse intensities that are comparable to those achieved under one-photon excitation. This finding opens the way to use both genetically targeted and pharmacologically selective azobenzene photoswitches to dissect intact neuronal circuits in three dimensions.