Mice can be conditioned to attend to one sensory modality while ignoring a second modality.

a, Behavioral setup. Mice were head-fixed and presented with a visual stimulus (a drifting grating on a screen) or a tactile stimulus (an air puff to the distal portion of the whiskers). b, Schematic for the shaping version of the task, depicting the timing of the air puff (red), the drifting grating (dark blue), and the water reward (light blue). Mice were presented with either stimulus with an inter-stimulus-interval of 8-12 seconds. In the tactile cohort, the air puff was paired with a water reward; in the visual cohort the drifting grating was paired with the water reward. c, Schematic for the full version of the task. Stimuli were allowed to overlap so that only the conditioned stimulus predicted the timing of the water reward. d, Timing of stimuli in the full task. Inter-stimulus intervals were drawn from an exponential distribution with a linear offset (top). As a result, the hazard rate was largely flat for most stimulus presentations (bottom). e, Example behavior from a mouse trained on the visual task. Top row: raster plot of licks aligned to the onset of the grating (blue region). Red markers indicate the first lick to occur after reward was available. Middle row: raster plot of licks aligned to the onset of the air puff (red region). Bottom row: Mean lick rate aligned to the onset of the drifting grating (blue) or the air puff (red). Gray region indicates the stimulation period. Data are from the first session of the shaping task (left column), the first day of the full task (middle column), and the final day of conditioning (right column). f, Learning curve of the mouse shown in E, showing the lick index (mean +/- SEM) in response to the drifting grating (blue) or the air puff (red) on each training day. Vertical dotted line indicates the day the mouse was switched from the shaping version to the full version of conditioning. Triangles: sessions shown in e. g, h, Similar to e and f but for an example mouse trained on the tactile task. i, Learning curves of all visually conditioned mice (left) and tactilely conditioned mice (right). Thin lines: mean lick index of a single mouse. Thick lines: mean lick index across all mice. j, Mean lick index for all mice on the first day of conditioning (“shaping session 1”), the first day of the full task (“full session 1”), and the final conditioning session (“final full session”).

Array recordings were made simultaneously from POm and LP.

a, Example coronal section. Red: DiI marking recording location. Gray: endogenous biotin stained with Alexa fluor 647 streptavidin. b, Zoom of the outlined region in a. The borders of LP, POm, and VPM are outlined. A 64-channel silicon probe was used to record from LP and POm. c, Mean waveform of an example POm cell across all probe channels. The channel with the largest mean waveform was identified and used to estimate the position of the cell (red circle in B). d, e, f, Estimated location of putative cells (gray circles) located in POm (red) and LP (blue) aligned to the Allen reference atlas. Cell locations are jittered along the medial-lateral and anterior-posterior axes for visualization purposes. d, Coronal projection over the range of cell locations in the anterior-posterior axis. e, Sagittal projection over range of cell locations in the medial-lateral axis. f, Transverse projection over range of cell locations in the dorsal-ventral axis.

POm responses to tactile and visual stimuli depend on conditioning paradigm.

a, Example cells from a tactilely conditioned mouse (same mouse as in figure 1e, conditioning session 7). Top row: spike raster aligned to the onset of the drifting grating (blue). Middle row: spike raster aligned to the air puff (red region). Bottom row: Mean firing rate over each trial aligned to the drifting grating (blue line) and the air puff (red line). Gray region indicates the timing of both stimuli. b, Sample cells from a visually conditioned mouse (same mouse as in figure 1c, conditioning session 12). c, Firing rate of all POm cells from tactilely conditioned mice aligned to either the air puff (left) or the drifting grating (right). Cells are sorted by the timing of the peak firing rate when aligned to the air puff. n=347 cells, 11 mice, range=11-57 cells/mouse, median=33 cells/mouse. d, Scatter plot of POm firing rates 1-second pre-stimulus (x-axis) and 1 second during stimulus onset (y-axis) from tactilely conditioned mice. Colored markers: cells with significantly different firing rates pre- and post-stimulus (1-way ANOVA with Holm-Bonferroni correction and post-hoc Wilcoxon signed-rank test). Gray markers: cells without significantly different firing rates. e, Firing rates of all POm cells from visually conditioned mice aligned to either the air puff (left) or the drifting grating (right). Cells are sorted by the timing of the peak firing rate aligned to the grating. (n=257 cells, 11 mice, range=10-42 cells/mouse, median=19 cells/mouse). f, Scatter plot of POm firing rates pre- and post-stimulus onset for visually conditioned mice, as in e. g, Event-triggered average firing rates of all POm cells (mean +/-SEM) aligned to either the air puff (red) or the drifting grating (blue) in tactilely conditioned mice (left) and visually conditioned mice (right). Gray region indicates the timing of both stimuli. h, Box plots of the change in firing rate from baseline for each POm cell during the first second of stimulus onset (“stimulus period”) and during the first two seconds post-stimulus (“offset period”). In tactilely conditioned mice, firing rates were higher in the air puff offset period than the drifting grating offset period (two-way ANOVA with post-hoc Wilcoxon signed-rank test. Conditioning type F=2.06, p=0.15; stimulus type F= 138, p<0.001; interaction F=133, p<0.001; tactile conditioning Wilcoxon p<0.001; visual conditioning Wilcoxon p=0.58). In both conditioning types, firing rates in the offset period were significantly different between stimulus types (conditioning F=0.12, p=0.73; stimulus F=16, p<0.001; interaction F=304, p<0.001; Wilcoxon p<0.001 for both visually and tactilely conditioned mice).

POm responses to tactile and visual stimuli vary based on anatomical location.

a, Sensory response index for significantly responding POm cells (colored cells in 3d and 3f). Cells with a positive selectivity index primarily respond to the air puff and cells with a negative index primarily respond to the drifting grating. Cells from tactilely conditioned mice had a significantly higher response index (Wilcoxon rank sum test, p<0.001, n=200 tactile conditioning cells, 120 visual conditioning cells). b, Position of all POm cells separated by conditioning type. Cell positions are projected on the coronal plane (top row) and sagittal plane (bottom row), as in figure 2d and 2e. Significantly responding cells are colored by their response index. Gray Xs indicate nonresponding cells. Positions in the medial-lateral and anterior-posterior axes are jittered for visualization only. c, Response index of significantly responding cells as a function of their position in POm along the anterior-posterior (left), dorsal-ventral (middle), and medial-lateral axes (right). Light gray: tactile conditioning. Dark gray: visual conditioning. A linear model of response index with experiment type and anatomical position as predictors revealed a significant effect of medial-lateral position and an interaction between position and experiment type. In visually conditioned mice, cells in the lateral portion of POm were more selective to the air puff (model p<10-60, adjusted R2=0.61, conditioning p<10-5, medial-lateral p<10-4, ML-conditioning interaction p<10-4, all other terms p>0.1)

POm correlates with arousal and movement regardless of conditioning.

a, Pupil and whisker tracking. Top left: Example frame from an eye video with the edge of the pupil highlighted in green. Top Right: A sample trace of pupil radius over time (gray: raw; green: smoothed). Bottom Left: Example frame from a whisker video with whiskers highlighted in green. Bottom Right: Example trace of whisker angle over time (gray) and the most protracted and retracted whisker positions over each whisk cycle (green), which is used to compute whisking amplitude. b, Whisking amplitude and pupil radius aligned to stimuli in tactilely conditioned mice (n=11 mice, left column) and visually conditioned mice (n=11 mice, right column). Top row: Pupil radius of each mouse aligned to either the air puff (blue) or the drifting grating (red) (Z-scored, mean+/-SEM). Bottom row: Whisking amplitude of each mouse aligned to the two stimuli (mean+/-SEM). Gray regions indicate stimulus timing. c, Cross-correlations between POm activity and pupil radius (top), whisking amplitude (middle), and lick rate (bottom). Light gray: Cells from tactilely conditioned mice (mean +/-SEM, n=347 cells, 11 mice). Dark gray: Cells from visually conditioned mice (n=257, 11 mice). Negative lags indicate that POm activity precedes the movement variable and positive lag indicates that POm activity follows the movement variable. d, Movement-corrected firing rates. The firing rate of each cell was fit to a linear model with whisking, licking, pupil radius, and baseline rate as predictors. Model residuals of all cells (mean+/-SEM) are aligned to the drifting grating and the air puff, as in B. e, Box plot of the model residuals of each POm cell during the first second of stimulus onset (“stimulus period”), and during the first two seconds post-stimulus (“offset period”), the same time windows as in 3g and 3h. Adjusted firing rates in tactilely conditioned mice were significantly higher during the air puff stimulus period than the drifting grating period. (Two-way ANOVA with post-hoc Wilcoxon signed-rank test. Conditioning type F=4.23, p=0.04; stimulus type F= 68.2, p<0.001; interaction F=38.5, p<0.001; tactile conditioning Wilcoxon p<0.001; visual conditioning Wilcoxon p=0.09). In visually conditioned mice, adjusted firing rates were higher during the drifting grating offset period than the air puff offset period. (conditioning F=0.003, p=0.95; stimulus F=3.9, p=0.048; interaction F=9.45, p=0.002; tactile conditioning Wilcoxon p=0.94, visual conditioning p<0.001).

Conditioning also reshapes LP activity.

a, Baseline firing rates of all POm and LP cells separated by conditioning type. POm cells had higher firing rates than LP cells, and firing rates did not significantly differ by conditioning type (two-way ANOVA, region F=8.0, p=0.0049; conditioning type F=0.44, p=0.51; interaction F=1.44, p=0.23). b, Example LP cells from a tactilely conditioned mouse (top) and a visually conditioned mouse (bottom). Mean firing rates over each trial are aligned to the drifting grating (blue line) and the air puff (red line). Gray region indicates the timing of both stimuli. c, Firing rate of all LP cells from tactilely conditioned mice aligned to either the air puff (left) or the drifting grating (right). Cells are sorted by the timing of the peak firing rate when aligned to the air puff. n=64 cells, 7 mice, range=4-16 cells/mouse, median=7 cells/mouse. d, Firing rate of all LP cells from visually conditioned mice aligned to either the air puff (left) or the drifting grating (right). Cells are sorted by the timing of the peak firing rate when aligned to the drifting grating. n=67 cells, 8 mice, range=1-19 cells/mouse, median=6 cells/mouse. e, Event-triggered average firing rates of all LP cells (mean +/SEM) aligned to either the air puff (red) or the drifting grating (blue) in tactilely conditioned mice (left) and visually conditioned mice (right). Gray region indicates the timing of both stimuli. f, Movement-corrected firing rates. The firing rate of each cell was fit to a linear model with whisking, licking, pupil radius, and baseline firing rate as predictors. Model residuals of all cells (mean+/-SEM) are aligned to the drifting grating and the air puff, as in E.

Manual curation of POm and LP cells

a, b, Example POm cells. Left, Mean spike waveform (black) with 200 randomly sampled individual waveforms (gray). Right, Autocorrelogram, binned at 0.5ms. Gray lines indicate a time lag of +/-1ms. c, d, Example LP cells, as in a and b. e, Mean waveforms of 50 randomly selected POm cells (red) and 50 randomly selected LP cells (blue). f, Histogram of waveform half-widths of each recorded POm cell (red, n=604) and LP cell (blue, n=131). g, Box chart of waveform half-width. Half-widths were not significantly different between LP and POm cells (POm: 0.40+/-0.07ms, LP: 0.41+/-0.08ms (mean+/-std), t-test p=0.052). h, Histogram of waveform amplitudes from each recorded POm and LP cell. i, Box chart of waveform amplitudes. POm cells had significantly higher amplitudes than LP cells (POm: 0.91+/-0.47mV, LP: 0.67+/-0.33mV (mean+/-std), t-test p<0.001).

Conditioning alters sensory response latency in POm and LP

a, b, Sample sensory response measurements of a POm cell from a visually conditioned mouse (a) and a LP cell from a tactilely conditioned mouse (b). A cell’s response latency was measured as the first time the mean firing rate exceeded a 99% confidence interval for at least two 10 ms time bins. Red: firing rate aligned to air puff onset. Blue: firing rate aligned to drifting grating onset. Dotted gray line: 99% confidence interval of baseline firing rate. Black triangle: response latency. Cells that did not respond to a stimulus within 500 ms were excluded from further analysis. c, Box chart of POm response latency to the air puff after tactile conditioning (gray) and visual conditioning (white). The air puff response latency was shorter in visually conditioned mice (n=219 responding cells from tactilely conditioned mice, 130 from visually conditioned mice; p<0.005, Wilcoxon rank-sum test). d, LP response latency to the air puff. There was no difference in response latency to the air puff between tactilely and visually conditioned mice (n=57 responding cells from tactilely conditioned mice, 46 from visually conditioned mice; p=0.103). e, LP response latency to the drifting grating. Response latency was longer in visually conditioned mice (n=27 responding cells from tactilely conditioned mice, 25 from visually conditioned mice; p<0.001).

LP responses to tactile and visual stimuli as a function of anatomical location

a, Sensory response index for significantly responding LP cells. Cells with a positive selectivity index primarily respond to the air puff and cells with a negative index primarily respond to the drifting grating. Cells from tactilely conditioned mice had a significantly higher response index (Wilcoxon rank sum test, p<0.001, n=50 tactile conditioning cells, 41 visual conditioning cells). b, Position of all LP cells separated by conditioning type. Cell positions are projected on the coronal plane (top row) and sagittal plane (bottom row), as in figure 2d and 2e. Significantly responding cells are colored by their response index. Gray Xs indicate nonresponding cells. Positions in the medial-lateral and anterior-posterior axes are jittered for visualization only.