Extended Data Fig. 3: Different types of movements recruit distinct but overlapping populations of spatially proximal dSPNs and iSPNs.

a, To study and compare movements of different types made by freely moving mice, we used custom software to extract from the behavioural videos the intervals from −4 to 4 s surrounding the onset of each movement bout. Using this software, we manually labelled each bout as an instance of forward locomotion, a left or right turn, grooming or upward rearing. If the mouse made multiple types of movement within an individual bout, we labelled the bout according to the first movement type exhibited, as only the interval from −1 to 2 s relative to motion onset was used in subsequent analyses of the accompanying neural Ca²⁺ activity. b, The fraction (mean ± s.e.m.) of SPNs that exhibited Ca²⁺ activity, relative to the baseline periods immediately preceding each movement type. Relative to baseline (dashed line), there was a significant increase in the fraction of dSPNs and iSPNs activated for all movement types except grooming. **P < 5 × 10⁻³ and ***P < 5 × 10⁻⁸ for dSPNs; ##P < 10⁻⁴ and ###P < 10⁻¹⁰ for iSPNs; Wilcoxon signed-rank test. Data in b and c are from n = 492 forward movements, 657 right turns, 810 left turns, 732 grooming and 204 rearing bouts from 17 Drd1a^cre mice, and n = 790 forward, 785 right turns, 1,015 left turns, 792 grooming and 164 rearing bouts from 21 Adora2a^cre mice. c, Rates (mean ± s.e.m.) of Ca²⁺ events in dSPNs and iSPNs, plotted as a function of time relative to the onsets of different types of movements. Event rates are shown normalized to the values of −2 to −1 s before motion onset and rose significantly above baseline values during all types of movement in both cell types. P < 10⁻⁶ for both cell types and all movement types; Wilcoxon signed-rank test. d, Mean values of the neural ensemble similarity computed for the sets of dSPNs and iSPNs that were active during pairs of bouts of either the same (on-diagonal) or different (off-diagonal) types of movements (Methods). For all movement types, the similarities of the cell ensembles that were active on different bouts of the same movement type were significantly greater than those of the ensembles that were active during the baseline periods before each bout. #P < 0.05; Kolmogorov–Smirnov test, corrected for multiple comparisons using a Benjamini–Hochberg procedure with a false-discovery rate of 0.05. Off-diagonal asterisks indicate that the neural ensembles that were active during bouts of two different movement types were significantly less similar to each other than the ensembles activated on different bouts of the same movement type, for both of the two movement types under consideration. *P < 0.05; Kolmogorov–Smirnov test, corrected for multiple comparisons using a Benjamini–Hochberg procedure with a false-discovery rate of 0.05. e, The cumulative distribution functions show the range of ensemble similarity values for the dSPN and iSPN ensembles that were activated on two bouts of the same movement type or on two bouts of different movement types (as described in d). For both iSPNs and dSPNs, ensemble similarity values were significantly lower for two bouts of different movement types than for two different bouts of the same movement type. P < 0.01; Kolmogorov–Smirnov test. f, Mean pairwise distances between the individual dSPNs or iSPNs activated on bouts of two different movement types. We normalized the values by comparing the mean weighted distance between the cells that were active during the two movement types to the same quantity determined under the null hypothesis that the spatial probability distributions of active cells on the two movement types were the same. For the latter determination, we created shuffled datasets in which we randomly permuted the firing rate of each cell between the two movement types, and we averaged the results over twenty-five different shuffled datasets. We normalized each distance value by taking the actual mean value, subtracting the mean value determined under the null hypothesis, and then dividing this difference by the standard deviation of the distance across the twenty-five shuffled datasets (Methods). Asterisks indicate that the mean pairwise distance was significantly greater than that expected by chance, indicating that the two movement types activated spatially distinguishable cell ensembles. *P < 0.05; Wilcoxon signed-rank test, corrected for multiple comparisons using a Benjamini–Hochberg procedure with a false-discovery rate of 0.05. Plots in d–f are based on n = 102 and n = 126 comparisons between bouts of the same movement type, and 255 and 315 comparisons of bouts of different movement types, in Drd1a^cre and Adora2a^cre mice, respectively. Values are from n = 17 Drd1a^cre mice and n = 21 Adora2a^cre mice, aggregated over the 1-h recordings on day −5 and the 30-min recordings performed on days −4 to −1 after saline vehicle injection but before drug administration.