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. 2024 Jul 15;27(9):1844–1857. doi: 10.1038/s41593-024-01697-1

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, Schematics shows imaging of striatal acute brain slices and photo-uncaging CYD8 with a 355-nm laser. b, Time-lapse images (semitransparent gray circle shows the field of UV illumination). Scale bar, 50 μm. c, Response of κLight1.3 to CYD8 photo-uncaging (blue, n = 6 slices) in the absence and presence of naloxone (Nalo, 10 μM; black, n = 3 slices). Solid lines represent the mean, and the shaded areas represent the s.e.m. d, Quantification of the peak ΔF/F evoked by CYD8 photo-uncaging. κLight1.3 (blue); 11.1% ± 1.36%, n = 6, + naloxone (black); 0.51% ± 0.12%, n = 3, P = 0.0011, two-tailed unpaired t-test. e, Time course of κLight1.2a after CYD8 (5 μM) photo-uncaging in the dStr. The dashed circle indicates the site of UV illumination. Heat map indicates ΔF/F (%). Scale bar, 50 µm. f, Summary of experiments determining the apparent diffusion coefficient, n = 7 slices from 4 mice. D* = 1.439 ± 0.37 μm² s⁻¹. g, Schematics show local electrical stimulation of hippocampal slice with trains of 1-s, 50-Hz stimuli with a 0.5-s interstimulus interval. HPC, hippocampus; ISI, interstimulation interval. h, Representative image showing expression of κLight1.3a in CA3 (top; scale bar, 0.5 mm) and zoomed in to visualize the localization of localization κLight1.3a to the membranes of neuronal processes in the dentate gyrus (middle; scale bar, 50 μm). Representative two-photon field of view from 15 stimulations (bottom) indicating the averaged intensity across all frames and z-score of responses in the representative field of view; scale bar, 20 μm. i,j, Average κLight1.3a responses to various electrical stimulation in the absence (i) and presence (j) of antagonists, Nor-BNI (gray) and ICI 174864 (green). Solid lines represent the mean, and shaded areas represent the s.e.m. k, Bar graph summarizing the peak fluorescence response to each stimulation condition. 15 stim (n = 8 slices): 14.3% ± 2.4%, 10 stim (n = 7 slices): 6.62% ± 0.8%, 5 stim (n = 7 slices): 4.28% ± 0.6%, 1 stim (n = 7 slices): 2.12% ± 0.3%, Nor-BNI (100 µM, n = 3 slices): 1.57% ± 1.2%, ICI 174864 (100 µM, n = 3 slices): 6.44% ± 0.3%. Ordinary one-way ANOVA with Bonferroni’s multiple-comparisons test, individual conditions compared to 15 stim, 15 stim versus 10 stim: **P = 0.044, 15 stim versus 5 stim: ***P = 0.0001, 15 stim versus 1 stim: ****P < 0.0001, 15 stim versus Nor-BNI: ***P = 0.0002, 15 stim versus ICI 174864: not significant (NS), P = 0.0525. Error bars represent the s.e.m.