Fig 1.
Location-recovered recordings of single VPM and POm neurons during active and passive whisker deflections in behaving mice.
(A) Experimental paradigm: single whisker deflection by focal air puffs (teal) or by active touches of a pole (yellow). Neuronal responses in VPM or POm are recorded juxtasomally. (B) Example band-pass filtered juxtasomal recording of a VPM neuron (black, same neuron as in E) and whisker angle (orange) during air puffs (teal) and active touches (yellow). (C) Example labeled neuron in POm. Scale bar = 200 µm. (D) Reconstructed locations of recorded VPM (blue) and POm (purple) neurons; large dots indicate neurons with significant air puff responses (n = 16 animals, VPM = 24/43, POm = 14/26, responsive/total recorded neurons; comparison of 50 ms windows before and after air puff, p < 0.05 one-sided Wilcoxon signed-rank test). (E) Raster plots (top, 50 trials) and PSTHs (bottom) of representative single neuron responses to puff in VPM (left) and POm (right). Mean whisker angle in orange, SEM indicated as shaded area. (F) Same as E but for active touch (yellow) responses. Shading indicates variable touch offset. Data and code underlying this figure can be found here: https://doi.org/10.5281/zenodo.14691035.
Fig 2.
POm shows pronounced response disparity to active and passive whisker stimuli.
(A) VPM (left) and POm (right) responses to air puffs (“Puff”, teal). Top: heatmaps of individual neuron PSTHs, normalized to baseline before puff and ordered by peak response latency. Bottom: mean population PSTHs, mean whisker angles (orange lines), and mean whisking amplitude over the plotted time range (−25 to 75 ms, orange bars) from recordings shown in the top panel. (B) Same as A but for active touches (“Touch”, yellow). (C) VPM spike rates during baseline and after whisker deflection via Puff or Touch. Baseline: 50 ms windows before, Puff/Touch: 50 ms after deflection. Individual neurons (black: rate increase p < 0.05 (9/15 neurons), dark gray: non-significant (6/15 neurons)) and population means (blue). Inset: pie chart of neurons with positive, negative, and non-significant responses to active touches. (D) Same as C but for POm (purple). Rate increase: 4/11 neurons, rate decrease: 1/11 neurons (light gray), non-significant: 6/11 neurons. (E) Modulation of Puff and Touch spike rates in comparison to baseline. Individual neurons (black) and population means for VPM (blue) and POm (purple), zero modulation indicated by red-dashed line. Asterisks represent p-values (*p < 0.05, **p < 0.01, ***p < 0.001); 2C–2E between conditions: two-sided Wilcoxon signed-rank test; 2E between neuron populations: two-sided Wilcoxon ranked-sum test; Individual neuron comparison (2C, 2D): one-sided (Puff) or two-sided (Touch) Wilcoxon signed-rank test; exact p-values, N numbers in S1 Table. Data and code underlying this figure can be found here: https://doi.org/10.5281/zenodo.14691035. Source data for panels C, D, E in S1 Data.
Fig 3.
Influence of whisking on whisker deflection responses in VPM and POm.
(A) Spike rates during quiescence (Q = no whisking) and during whisking (W) for individual neurons (black: whisking rate significantly different from base rate (p < 0.05, shuffle test, see Methods), gray: non-significant change) and population means (VPM: blue line, POm: purple line). (B) Spike rates as a function of whisking amplitudes for VPM (left) and POm (right), individual neurons (black: significant correlation p < 0.05, gray: non-significant increase, based on Student T cumulative distribution function), averages in blue and purple for VPM and POm, respectively. (C) Spike rates as a function of phase in the whisk cycle (gray bars) and a sinusoidal fit to the data (blue and purple for VPM and POm example neurons, respectively). (D) Polar plot of phase signal to noise (SNR, radius) and preferred phase (angle) for VPM (blue) and POm (purple) neurons. Filled circles, neurons with significant phase modulation (p < 0.05, Kuiper test). Phase zero refers to a fully protracted position. (E) Responses to air puffs (teal) during quiescence (Q-Puff) in VPM (left) and POm (right), PSTHs of individual neurons (heat maps, normalized to baseline before air puff and ordered by peak response latency) and population PSTHs. Mean whisker angles (orange lines) and mean whisking amplitudes (orange bars). (F) Same as E but for air puffs during whisking (W-Puff). (G) VPM spike rates during baseline and in response to air puffs when the animal was quiescent (Q) or whisking (W) Baseline: 50 ms windows before, Puff: 50 ms after. Individual neurons (black: rate increase p < 0.05, gray: non-significant increase) and population means (blue). (H) same as G but for POm (purple). (I) Modulation index of Q-Puff and W-Puff spike rates in comparison to baseline. Individual neurons (black) and population means for VPM (blue) and POm (purple), zero modulation indicated by red-dashed line. (J) Modulation index of W-Puff and Touch spike rates in comparison to baseline. Individual neurons (black) and population means for VPM (blue) and POm (purple), zero modulation indicated by red-dashed line. Asterisks represent p-values (*p < 0.05, **p < 0.01, ***p < 0.001); 3G–3J between conditions: two-sided Wilcoxon signed-rank test; 3I–3J between neuron populations: two-sided Wilcoxon ranked-sum test; Individual neuron comparison (3A, 3G, 3H): one-sided Wilcoxon signed-rank test; exact p-values, N numbers in S1 Table. Data and code underlying this figure can be found here: https://doi.org/10.5281/zenodo.14691035. Source data for panels A, G, H, I, J in S2 Data.
Fig 4.
Dependence of thalamic responses to active and passive whisker deflections on whisker kinematics and intervals.
(A) Distribution of absolute angular acceleration during (±25 ms) touch (yellow) and puff (teal). Joint distribution divided into acceleration tertiles (gray-shaded bars). (B) Modulation in response to puff (teal) and touch (yellow) split by acceleration tertiles indicated in A for VPM (left) and POm (right). (C) Smoothed VPM (left) and POm (right) population mean PSTHs split by tertiles (shades of gray) indicated in A for puff (top) and touch stimuli (bottom). (D–F) Same as in A–C but splitting deflection events into tertiles of whisker curvature following whisker deflection (0–25 ms). (G–I) Same as in A–C but splitting deflection events into tertiles based on intervals between whisker deflections. (J) Responses to 1st vs. subsequent touches within a bout. Individual neurons (black) and population means for VPM (blue, left, n = 10) and POm (purple, right, n = 10). Baseline: 50 ms windows before 1st touch in bout, Touch: 50 ms after. Only neurons with at least 10 touches in each condition were included. Asterisks represent significance (*p < 0.05); 4B, 4E, 4H within condition: one-sided Wilcoxon signed-rank, between tertiles and conditions: two-sided Wilcoxon signed-rank test; exact p-values in S2 Table. Data and code underlying this figure can be found here: https://doi.org/10.5281/zenodo.14691035. Source data for panels B, E, H, J in S3 Data.
Fig 5.
Inhibition of the barrel cortex (BC) increases VPM and POm correlation with free whisking but strongly attenuates POm whisker deflection responses.
(A) Experimental setup. BC was optogenetically suppressed while juxta-cellular recordings were made from single neurons in VPM and POm. A single whisker was deflected with air puffs. (B) Spike rates during quiescence (Q = no whisking) and during whisking (W) during BC suppression (Laser on) and control conditions (Laser off). Individual neurons (black lines: whisking rate significantly different from base rate (p < 0.05, shuffle test, see Methods), gray: non-significant change) and population means (VPM blue line, POm purple line). (C) Polar plot of phase-SNR (radius) and preferred phase (angle) for VPM (left, blue) and POm (right, purple) neurons, during control condition (Laser Off, circles) and during BC inactivation (Laser On, diamonds). Filled markers indicate significantly phase-modulated neurons per condition. Phase zero refers to a fully protracted position. (D) VPM (left) and POm (right) phase-SNRs when the laser was on or off. Individual neurons (black lines: significant phase modulation (p < 0.05, Kuiper test) during laser on condition, gray lines: nonsignificant phase modulation) and population means (VPM blue line, POm purple line). Percentage of neurons significantly modulated in laser on/off conditions. (E) VPM (left) and POm (right) responses to air puffs (teal) in control condition (Laser off). Top: heatmaps of individual neuron PSTHs normalized to baseline before air puff and ordered by peak response latency. Bottom: mean population PSTHs and whisker angles (orange). (F) Same neurons as in E but when the BC was inactivated (Laser on, light blue). (G) VPM spike rates during baseline and in response to air puffs during control conditions (Laser off) and during BC inactivation (Laser on). Baseline: 50 ms windows before, Puff: 50 ms after. Individual neurons (black lines: rate increase p < 0.05, gray lines: non-significant increase) and population means (blue lines). (H) same as G but for POm. (I) Rate modulation index in comparison to baseline for puff-evoked responses with and without BC inactivation. Individual neurons (black) and population means for VPM (blue) and POm (purple), zero modulation indicated by red-dashed line. Asterisks represent p-values (*p < 0.05, **p < 0.01, ***p < 0.001); 5G–5H, 5I between conditions: two-sided Wilcoxon signed-rank test; 5I between neuron populations: two-sided Wilcoxon ranked-sum test; Individual neuron comparison (5B, 5D, 5G, 5H): two-sided Wilcoxon signed-rank test; exact p values, N numbers in S1 Table. Data and code underlying this figure can be found here: https://doi.org/10.5281/zenodo.14691035. Source data for panels B, D, G, H, I in S4 Data.