High performance microbial opsins for spatially and temporally precise perturbations of large neuronal networks
High performance microbial opsins for spatially and temporally precise perturbations of large neuronal networks
Summary Patterned optogenetic activation of defined neuronal populations in the intact brain can reveal fundamental aspects of the neural codes of perception and behavior. The biophysical properties of existing optogenetic tools, however, constrain the scale, speed, and fidelity of precise optical control. Here we use structure-guided mutagenesis to engineer opsins that exhibit very high potency while retaining fast kinetics. These new opsins enable large-scale, temporally and spatially precise control of population neural activity in vivo and in vitro. We benchmark these new opsins against existing optogenetics tools with whole-cell electrophysiology and all-optical physiology and provide a detailed biophysical characterization of a diverse family of microbial opsins under two-photon illumination. This establishes a toolkit and a resource for matching the optimal opsin to the goals and constraints of patterned optogenetics experiments. Finally, by combining these new opsins with optimized procedures for cell-specific holographic photo-stimulation, we demonstrate the simultaneous co-activation of several hundred spatially defined neurons with a single hologram, and nearly double that number by temporally interleaving holograms at fast rates. These newly engineered opsins substantially extend the capabilities of patterned illumination optogenetic paradigms for addressing neural circuits and behavior.
Hendricks William D.、Ant¨?n Oldenburg Ian、Gajowa Marta、Ogando Mora B.、Jagadisan Uday、Sadahiro Masato、Bounds Hayley、Gopakumar Karthika、Tayler Ian、Abdeladim Lamiae、Brohawn Stephen G.、Sridharan Savitha、Adesnik Hillel
Department of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaThe Helen Wills Neuroscience InstituteDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of California||The Helen Wills Neuroscience InstituteDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of California||The Helen Wills Neuroscience Institute
生物科学研究方法、生物科学研究技术神经病学、精神病学分子生物学
Hendricks William D.,Ant¨?n Oldenburg Ian,Gajowa Marta,Ogando Mora B.,Jagadisan Uday,Sadahiro Masato,Bounds Hayley,Gopakumar Karthika,Tayler Ian,Abdeladim Lamiae,Brohawn Stephen G.,Sridharan Savitha,Adesnik Hillel.High performance microbial opsins for spatially and temporally precise perturbations of large neuronal networks[EB/OL].(2025-03-28)[2025-06-17].https://www.biorxiv.org/content/10.1101/2021.04.01.438134.点此复制
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