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. 2023 Oct 23;14:6712. doi: 10.1038/s41467-023-42288-1

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© The Author(s) 2023

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. 8 — A Left: Optogenetic stimulation of direct pathway striatal projection neuron (dSPN) axons in the globus pallidus externus (GPe) using the opsin ChrimsonR; simultaneous recording of Npas1 activity using the calcium indicator GCaMP6s. dStr dorsal striatum. Right: All-optical setup. B ChrimsonR-TdTomato+ dSPN terminals in the GPe (red) apposed to (white arrows) GCaMP6s+Npas1 somas (cyan). Optic fibers in the same region. Representative from N = 6 mice. C Opto-stimulation is triggered in a closed-loop when Npas1 dFF surpasses a defined threshold (see Methods). D 10 s, 20 Hz stimulation of dSPN GPe axons leads to a power-dependent reduction in Npas1 activity. Left: Average traces, Middle: Heatmaps of all trials, Right: Amplitude change in Npas1 dFF in the opto-window (ANOVA: virus x power p < 0.001; Sidak post-hoc: Chrimson **p < 0.01, ***<0.001, mCherry p > 0.8). E As expected: no effect of unilateral stimulation on mouse speed (ANOVA: power p = 0.50). F 10 s, 2 mW stimulation of dSPN GPe axons leads to a frequency-dependent reduction in Npas1 activity. Left: Average traces, Middle: Heatmaps of all trials, Right: Amplitude change in Npas1 dFF in the opto-window (ANOVA: virus x power p < 0.01; Sidak post-hocs: Chrimson: 0 vs 20 Hz *p < 0.05, 0 vs 10 Hz p = 0.99, mCherry p = 0.28 and 0.51). G No effect of stimulation on mouse speed (ANOVA: power p = 0.56). H Opto-stimulation triggered in a closed-loop during ongoing locomotion when mouse speed reaches a defined threshold (see Methods). I As expected, mouse speed increases before the opto-trigger (baseline vs. threshold (thresh) and stim periods), but is not affected by opto-stimulation (0 vs. 0.2 mW) (ANOVA: epoch p < 0.001, epoch x LED p = 0.79; Sidak post-hocs all ***p < 0.001). J 5 s, 20 Hz stimulation of dSPN GPe axons at ultra-low power (0.2 mW) reduces motor-related Npas1 activity. Left: Average and summary data showing a significant increase in dFF before the opto-trigger (ANOVA: epoch p < 0.05). Middle: Heatmaps of all trials, Right: Amplitude change in Npas1 dFF in the opto-window (ANOVA: virus x LED p < 0.01; Sidak post-hocs: Chrimson **p < 0.01, mCherry p = 0.84). Heatmaps: straight/dashed line = LED onset/offset. N = 5 mCherry, N = 6 ChrimsonR throughout. Data are mean ± SEM. Exact p-values are given in Supplementary Dataset S2. See also Supplementary Fig. S9. Source data are provided as a Source Data file.