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. 2021 Jul 23;12:4488. doi: 10.1038/s41467-021-24718-0

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© The Author(s) 2021

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Example recordings of two neurons (10 s and 1 s; top). Neurons were held in current clamp (pre) and stimulated with 470 nm blue light (blue box). Light stimulation increased the firing rate of all firing Opn7b-expressing neurons (10 s: n = 13 cells from 11 animals and 1 s: n = 8 cells from 11 animals; bottom). Time constant of activation (τ_light) of light-induced maximum firing rate is shown in red. b Mean response rate of Opn7b-expressing cells increased by light-induced deactivation (deACT) of Opn7b with blue light (10 s: n = 13 cells from 11 animals; 1 s: n = 8 cells from 11 animals). In contrast, light stimulation did not change the firing rate of enhanced green fluorescent protein (eGFP)-expressing neurons (n = 5 from 2 animals). Mean values (±SEM) and single-cell data were plotted, significance was assessed using two-sided Wilcoxon signed-rank test. c Example trace of a neuronal response to a −100, 0, 100, and 200 pA pulse before stimulation (pre), direct after light-induced deACT of Opn7b and after a recovery time of at least 3 min after stimulation (recovery). d Comparisons of membrane potentials in Opn7b-expressing neurons before (pre, black, n = 30 from 12 animals) and after light stimulation (blue, deACT, n = 22 cells from 11 animals and gray, recovery, n = 12 cells from 5 animals) and eGFP-expressing neurons before and after light stimulation (green, eGFP: n = 17 cells from 2 animals, eGFP-stim: n = 12 cells from 2 animals). Mean values (± SEM) and single-cell data (circles) are shown. Significance was assessed using Kruskal-Wallis-test (p < 0.001) followed by pairwise Dunn’s test. e Comparisons of rheobase in Opn7b-expressing neurons before (pre, black, n = 30 cells from 12 animals) and after light stimulation (blue, deACT, n = 22 cells from 11 animals; and gray, recovery, n = 12 cells from 5 animals) and eGFP-expressing neurons before and after light stimulation (green, eGFP: n = 17 cells from 2 animals; eGFP-stim: n = 12 cells from 2 animals). Mean values (±SEM) and single-cell data (circles) are shown. Significance was assessed using Kruskal-Wallis-test (p = 0.099). f Example trace of a neuronal response to various small current steps from −100 to 100 pA pulse before stimulation (pre) and direct after deACT used to calculate the input resistance. g I-V plot generated from the voltage response before (black) and after light-induced deactivation (blue) shown in (f). The average input resistance was estimated from the slope of the I-V plot between −100 and −40 pA. h Mean input resistance of Opn7b-expressing cells increased by light-induced deACT with blue light (n = 24 cells from 11 animals). Mean values (±SEM) and single-cell data were plotted, significance was assessed using two-sided Wilcoxon signed-rank test. Source data are provided as a Source Data file.