Extended Data Fig. 6: Dopaminergic genetic subtypes show different signaling patterns during locomotion.
From: Unique functional responses differentially map onto genetic subtypes of dopamine neurons
(a) Locomotion response (PC1/PC2) mapped onto recording location for each subtype and DAT. Same as Fig. 3b but without collapsing slices for compactness and without random mediolateral shifting of recording locations to reduce overlap. (b) Average ΔF/F triggered on large accelerations (left, ▲) and large decelerations (right, ▽) for Aldh1a1 recordings (as Fig. 2g). Isosbestic control shown in light blue, same scale as ΔF/F average but shifted. Acceleration shown in grey in the background (scale bar = 0.2 m/s2). Shaded regions denote mean ± s.e.m. Heatmap shows triggered average for each recording, sorted by PC1/PC2 angle (see Fig. 2l). Aldh1a1 mice = 14, n = 75 recordings. (c) Average cross-correlation between ΔF/F traces and acceleration for Aldh1a1 recordings (as Fig. 2f). Isosbestic control shown in blue. Shaded regions denote mean ± s.e.m. Heatmap shows cross-correlation for each recording, sorted as in B. (d) Average acceleration triggered on large transients for Aldh1a1 recordings (as Fig. 2h). ΔF/F average and isosbestic control shown in the background (scale bar = 5% Norm ΔF/F.) Shaded regions denote mean ± s.e.m. Heatmap shows triggered average for each recording, sorted as in B. (e) Principal component scores for each recording along PC1 and PC2 for Aldh1a1 (same as Fig. 2k) but with each Aldh1a1 recording color-coded by depth within striatum, showing that Aldh1a1 axons deeper in striatum show similar locomotion signaling to Calb1. (f) Timing of the calcium transient peak in triggered averages on decelerations (Fig. 2g, right) for each recording from Calb1 and Vglut2. Means Vglut2 = 0.35, Calb1 = 0.23; p-value for comparison between subtypes = 0.01 (two-sided Wilcoxon rank-sum test). Vglut2 mice = 12, n = 42 recordings; Calb1 mice = 6, n = 22 (as in Fig. 2i). Error bars denote mean ± s.e.m. (g) Timing of the deceleration peak in triggered averages on ΔF/F transient peaks (Fig. 2h) for each recording from Calb1 and Vglut2. Means Vglut2 = 0.47, Calb1 = 0.34; p-value for comparison between subtypes = 0.005 (two-sided Wilcoxon rank-sum test). Same n as F. Error bars denote mean ± s.e.m. (h) The locomotion signaling observed in DAT mice across depths (H) can be explained by mixtures of the Anxa1 and Calb1 subtypes in varying ratios matching the relative abundance of each subtypes’ axons in that depth (H’). (i) Average cross-correlation between ΔF/F traces and acceleration for all recordings of each functionally homogeneous subtype (as Fig. 2f) but averaged per mouse. Shaded regions denote mean ± s.e.m. Heatmap shows cross-correlation average for each mouse, sorted by PC1/PC2 angle (see Fig. 2l). (j) Difference in locomotion signaling (measured as the difference in PC1/2 angle, as shown in Fig. 2l) between pairs of recordings made at difference distances from each other, for pairs of recordings from the same subtype (Vglut2, Calb1, and Anxa1, in colors), from DAT mice (mixture of subtypes, in grey), or from mismatched subtypes (Vglut2-Calb1, Vglut2-Anxa1, and Calb1-Anxa1). P-values for comparison between pairs within same subtype vs mismatch subtypes: Vglut2 = 3 × 10−05, 6 × 10−23, 2 × 10−25, 5 × 10−25, 1 × 10−08, 0.03, 9 × 10−04, 1, 0.9; Calb1 = 2 × 10−04, 5 × 10−22, 4 × 10−10, 1 × 10−07; Anxa1 = 8 × 10−08, 6 × 10−35, 1 × 10−21, 5 × 10−09, 0.06, 0.02, 1, 1 (two-sided Mann-Whitney U test with Bonferroni correction). Number of pairs of recordings per distance bin (from 0 in steps of 0.3 mm): Vglut2 = [24, 107, 108, 91, 58, 29, 28, 14, 6, 0, 0], Calb1 = [37, 83, 29, 18, 1, 3, 0, 0, 0, 0, 0], Anxa1 = [252, 410, 250, 52, 49, 30, 28, 10, 0, 0, 0], Mismatch = [47, 245, 438, 525, 461, 661, 542, 615, 367, 189, 49]. Error bars denote mean ± s.e.m.
