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. 2023 May 24;14:2999. doi: 10.1038/s41467-023-38798-7

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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 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. 5 — a stGtACR2-FusionRed-expression (red) in L5 S1HL of an Rbp4-Cre mouse showing fluorescence in L5 neurons, S1HL layer borders (dashed lines), estimated based on soma sizes and densities using DAPI signals (blue). A representative example from n = 15 mice. b Schematic of silicon-probe recordings in S1HL in a head-fixed awake mouse (upper); example raster demonstrating inhibition of spiking in an example L5 unit (lower). c Example raster for a laser-suppressed L5 stGtACR2 unit (depth = 589 μm). d Upper panel: depth-resolved light modulation indices (MI_L) for L2/3, L4, L5, and L6 units in S1HL during optogenetic inhibition of L5 (n = 3 pooled experiments, 285 units). Each unit is represented by one data point; the size and color of markers are proportional to the spontaneous spiking rate (black to yellow: low to high). Depth values are slightly jittered for visibility. Non-significantly modulated units are not shown (122/285). Lower panel: MI_L distributions for significantly modulated units by layer. e Within-animal comparison of paw withdrawal probabilities in response to graded von Frey stimulation of the hindpaw at baseline (black, laser off) and during optogenetic inhibition (red, laser on, 5 s continuous pulse) in the contralateral S1HL of L5-stGtACR2 mice (n = 6). f Conditioned place aversion (CPA) test. Population analysis of total time spent in the laser-paired chamber at baseline (Laser off, black/gray) and during inhibition (Laser on, red 20 Hz laser stimulation in S1HL cortex) of L5-EGFP (n = 7) and L5-stGtACR2 (n = 6, p = 0.006) naive mice. g Average chamber preference indices (PI) for L5-stGtACR2 (n = 6) and L5-EGFP (n = 7) mice. A PI of 1 indicates a full preference, while a PI of −1 indicates full avoidance of the laser-paired chamber. PIs were not significantly different between groups during laser stimulation or at baseline. * represent p < 0.05; 5d: two-sided Wilcoxon signed-rank test; see Supplementary Table 1 for population medians and first and third quartiles per layer and condition.; 5e–g: Two-way repeated measures ANOVA with post hoc Bonferroni test. Exact F and p values are in Supplementary Table 1. Data were shown as mean ± SEM. Source data for c–g are provided as a Source Data file.