Figures and data
Patchy SPN ablation reduces rest and unmasks anxious vigor at choice points.
(A) Injection schematic. (B) MOR1 staining. Scale bar: 500μm. (C) MOR1 quantification to assess ablation. 3 mice per group, n=22 Control (Ctrl) and 30 patchy SPN ablated (PA) hemisections. For dorsal striatum (DS) PA vs Ctrl 2-tailed Mann-Whitney****p<0.0001. (D-J) Light/Dark box. n=11 Ctrl and 10 PA mice. (D) LDbox schematic. (E) % time in dark. (F) Average speed. Following RM ANOVA, post hoc comparisons ***p=0.001, *p=0.0371; **p=0.0015 (G) Maximum speed. 2-tailed Mann-Whitney *p=0.0434. (Hi) Speed distribution normalized to zone. (Hii) %time ≤2cm/sec paired t-test L vs D Ctrl **p=0.001, PA p>0.05; unpaired t-test Ctrl vs. PA in dark **p=0.0018, in light p>0.05. (I) Transition speed. (J) GLME 95% C.I. for impact of BIL on transition speed. (K-O) LL/DD box. n=7 Ctrl-LL, 6 Ctrl-DD, 5 PA-LL, 6 PA-DD mice. (K) LL/DDbox schematics. (L) Average speed of LL/DDbox test. Mixed Effects analysis ns. (M) Maximum speed of LL/DDbox test. Mixed Effects analysis ns. (N) Speed distribution normalized to side “A” of LL or DD box. (O) transition speed. (P) GLME 95% C.I. for impact of BIL on transition speed overlaps zero for both groups.
Patchy SPN activity reflects zone, speed, deceleration.
Single cell calcium imaging with miniscopes. n=10 mice. (A) Schematic illustrating placement of GCaMP6s and GRIN lens. (B) Example lens placement above imaged patchy SPNs. Selection expanded at right shows GCaMP6s and MOR1 overlap. Scale bar: 500 μm. (C) Left: image stack collected through GRIN lens with location of example cells circled. Scale bar: 500 μm. Right: calcium transients from example cells circled at left plotted above speed. (D) Light/Dark box aerial view with tethered mouse. (Ei) Average activity of zone-preferring (Light or Dark) and non-discriminating (Other) neurons in light “L” vs dark “D” zones. paired t-test p<0.0001 Light cells; p<0.0001 Dark cells; p=0.016 Other cells. (Eii) Most zone-discriminating neurons are light-preferring. (Fi) Sample neurons illustrating speed relationships; clockwise from top left: linear+, linear-, quadratic+, quadratic-. (Fii) Distribution of speed relationships among imaged neurons. (G) Distribution is similar across all mice (n=9, excluding one mouse with fewer than 80 cells) for (Gi) zone and/or speed related neurons (Gii) specific speed relationships (H) Speed relationships among light- and dark-preferring neurons. (I) Histogram of ADI. Strong deceleration-predicting “DP” neurons are defined by |ADI| > a threshold “thr” of 0.8. (J) DP neurons are over-represented among zone/speed free neurons, χ2(1,1565) =24.33, p<0.0001.
Light sensitivity and speed modulation are reflected in transition-active neurons.
Single cell calcium imaging with miniscopes. n=10 mice. (A) A relationship exists between zone/speed encoding and zone transitions, χ2(4, n=1563) =137.3, p<0.0001. (B) A relationship exists between deceleration encoding and zone transitions, χ2(1, n=1565) =24.33, p<0.0001. (C) comparison of transition-active neurons selective for either BIL or BID. (D) Overlay of average ΔF/F (green, z-score) with mean transition speed (grey scale, cm/s). (E) effect of BIL while approaching or retreating from zone transitions, GLME fixed effects 95% CI (a.u.) (F) Mean R value per mouse for all acceleration events with significant (p<0.05) ΔF/F cross-correlation to (Fi) speed (n=9 into light, 9 into dark, 10 within light, 10 within dark), or (Fii) acceleration (n=3 into light, 5 into dark, 8 within light, 9 within dark).
Patchy SPN efferents to SNr encode speed and deceleration.
Sepw1-Cre: n=8 GPe, n=9 GPi, n=8 SNr; Calb1-Cre: n=8 SNr. 95% CI shown in red. (A) injection schematic for mice with later fiber implants to GPe, GPi, or SNr. (B) AAV1-phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP injected to dorsal striatum of Sepw1-Cre mouse illustrating patchy SPN efferents at GPe, GPi, and SNr (circled). Clockwise from top right: TH stain, tdTomato, eGFP, overlay. Scale bar 500µm. (C) Patchy SPN efferent activity aligned to zone transition, overlaid with speed. GCaMP8s into Light (light green) or Dark (dark green); simultaneous 405nm channel into Light (light blue) or Dark (dark blue); mean speed (cm/s) during transition into Light (grey) or Dark (black). (D) average across mice of mean R-value for acceleration events significantly correlated to ΔF/F (p<0.05) during zone transitions (E-L) Cross-correlogram color code: transition into Light (light green) or Dark (dark green), or within Light (light blue) or Dark (dark blue). (E-H) Patchy SPN ΔF/F at SNr cross-correlation to speed or acceleration aligned to acceleration events. (Ei) heat map of cross-correlation to speed for each mouse (Eii) ΔF/F cross-correlogram to speed. (Fi) heat map of cross-correlation to acceleration for each mouse (Fii) ΔF/F cross-correlogram to acceleration. (G-H) 95% CI in red. (G) for all acceleration events with significant (p<0.05) ΔF/F cross-correlation to speed, (Gi) mean R value per mouse, (Gii) mean Xcorr lag per mouse. (H) for all acceleration events with significant (p<0.05) ΔF/F cross-correlation to acceleration, (Hi) mean R value per mouse, (Hii) mean Xcorr lag per mouse. (I) Patchy SPN efferent ΔF/F at SNr inter event interval in either zone, paired ttest ***p=0.0001 (J-M) matrix ΔF/F at SNr (n=8) (J) matrix efferent activity at SNr aligned to zone transition, overlaid with speed. Color coding identical to C. (K) average across mice of mean R-value for acceleration events significantly correlated to ΔF/F (p<0.05) during zone transitions (Li) ΔF/F cross-correlogram to speed. (Lii) ΔF/F cross-correlogram to acceleration. (M) matrix efferent ΔF/F at SNr inter event interval in either zone, paired ttest **p=0.0039
Patchy SPN enhancement increases rest and eliminates discriminative speed at choice points.
(A) injection schematic. (B) Dorsal striatal DREADD-mCherry overlaps patch marker MOR1. Scale bar: 500μm. 20 × channel separation at right. (C) Dorsal striatal quantification of MOR1 and mCherry in Ctrl or Gq-DREADD-transduced mice. N=one dorsal striatal subregion (from Bregma in mm: R: 0.98-1.8, M: 0.5-0.7, C: 0.02-0.26) average value from n=4-6 sections per subregion, representing 3 Ctrl and 5 Gq mice. (D-I) Light/Dark box. n=17 Ctrl and 20 Gq mice. (D) % time in dark, p>0.05. (E) Average speed. Ctrl-L vs Ctrl-D, ***p=0.0001, Gq-L vs Gq-D, p=0.0037; Ctrl-vs-Gq in Light **p=0.0044, in Dark *p=0.0417. (F) Maximum speed. *p=0.01. (Gi) Speed distribution normalized to zone. (Gii) %time ≤2cm/sec L vs D Ctrl ****p<0.0001, Gq **p=0.0056; Ctrl vs Gq in Light **p=0.0059, in Dark *p=0.0168. (H-I) Transition speed for controls (Hi) and Gq (Hii). (I) 95% C.I. for GLME by group shows speed approaching transitions is increased by BIL for controls but reduced for Gq-mice. (J) Cartoon summarizing speed modulation by valence under control (blue) SA (red) and Gq (green) conditions.
Patch/matrix balance at SNr controls Light/Dark box locomotor phenotype.
(A) Injection schematic. (B) Sample image of TH (magenta) and mCherry (red) staining in SN. The fiber track is indicated with dashed line. Scale bars: 500μm. (C-H) n=14 Ctrl and 16 ChR Sepw1-Cre mice. (C) % time in dark, p>0.05. (D) Average speed. Ctrl-L vs Ctrl-D, ****p<0.0001, ChR-L vs ChR-D, **p=0.0011; Ctrl-vs-ChR in either zone p>0.05. (E) Maximum speed. p>0.05. (Fi) Speed distribution normalized to zone. (Fii) %time ≤2cm/sec L vs D Ctrl ***p=0.0002, ChR ***p=0.0005; Ctrl vs ChR in either zone p>0.05. (G-H) Transition speed for controls (Gi) and ChR (Gii). (H) 95% C.I. coefficient for GLME by group shows speed approaching transitions is increased by BIL for controls significantly more than for ChR mice. (I) Cartoon summarizing speed modulation by valence under control (blue) Gq (green) and SNr ChR (purple) conditions for SpCre mice. (J) injection schematic. (K) Sample image of TH (magenta) and mCherry (red) staining in SN. The fiber track is indicated with dashed line. Scale bars: 500μm. (L-Q) n=13 Ctrl and 13 ChR Sepw1-Cre mice. (L) % time in dark, p>0.05. (M) Average speed. Ctrl-L vs Ctrl-D, ****p<0.0001, ChR-L vs ChR-D, **p=0.0012; Ctrl-vs-ChR in either zone p>0.05. (N) Maximum speed. p>0.05. (Oi) Speed distribution normalized to zone. (Oii) %time ≤2cm/sec L vs D Ctrl ****p<0.0001, ChR p>0.05; Ctrl vs ChR in either zone p>0.05. (P-Q) Transition speed for controls (Pi) and ChR (Pii). (Q) 95% C.I. coefficient for GLME by group. (R) Cartoon summarizing speed modulation by valence under control (blue) SA (red) and Calb1-Cre SNr ChR (maroon) conditions.
