Patchy striatonigral neurons modulate locomotor vigor in response to environmental valence
- Transgenic Section, Laboratory of Neurogenetics, National Institute on Aging, National Institutes of Health, United States
- Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, United States
- School of Electrical Engineering & Computer Science, College of Engineering & Mines, University of North Dakota, United States
- The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, United States
Figures
Figure 1 with 1 supplement
Patchy spiny projection neuron (SPN) ablation reduces rest and unmasks anxious vigor at choice points.
(A) Injection schematic. (B) MOR1 staining. Scale bar: 500 μm. (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, two-tailed Mann-Whitney, ****p<0.0001. (D–J) Light/Dark box. n=11 (7Male [M],4Female[F]) Ctrl and 10 (6M,4F) 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. Two-tailed Mann-Whitney, *p=0.0434. (H) Speed distribution normalized to zone. Insert, %time ≤2 cm/s, 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) Generalized Linear Mixed-Effect (GLME) 95% C.I. for impact of body-in-light (BIL) on transition speed. (K–P) LL/DD box. n=7 (4M,3F) Ctrl-LL, 6 (3M,3F) Ctrl-DD, 5 (3M,2F) PA-LL, 6 (3M,3F) 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% CI for impact of BIL on transition speed overlaps zero for both groups.
Figure 1—figure supplement 1
Patchy distribution of mCherry-positive cells in the dorsal striatum of Sepw1-Cre mice.
Representative images show mCherry (red) and MOR1 (green) immunostaining in the dorsal striatum of Sepw1-Cre mice (n>3) following stereotactic injection of AAV-EF1a-DIO-mCherry. Scale bar: 500 mm.
Figure 2 with 1 supplement
Patchy spiny projection neuron (SPN) activity reflects zone, speed, deceleration.
Single-cell calcium imaging with miniscopes. n=10 mice (7 M,3F). (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. (E) 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. (F) Most zone-discriminating neurons are light-preferring. (G) Sample neurons illustrating speed relationships; clockwise from top left: linear+ (L+), linear- (L-), quadratic+ (Q+), quadratic- (Q-). (H) Distribution of speed relationships among imaged neurons. (I, J) Distribution is similar across all mice (n=9, excluding one mouse with fewer than 80 cells) for (I) zone and/or speed-related neurons (J) specific speed relationships. (K) Speed relationships among light- and dark-preferring neurons. (L) Histogram of acceleration/deceleration preference index (ADI). Strong deceleration-predicting ‘deceleration-prediction (DP)’ neurons are defined by |ADI|>a threshold ‘thr’ of 0.8. (M) DP neurons are over-represented among zone/speed free neurons, χ2(1,1565)=24.33, p<0.0001.
Figure 2—figure supplement 1
MOR1 thresholding, Scope-mounted Light/dark box behavior, and Ca2+ imaging supplemental.
(A) Patch ablation was assessed by MOR1-positive area measurements. Example dorsal striatal (DS) and ventral striatal (VS) hemisection stained for MOR1 (left) alongside thresholded image generated in MATLAB (right) used for quantifying MOR1-positive territories. N>3. Scale bar: 500 mm. (B) Example field of view through GRIN lens. Scale bar: 500 mm. (C) Schematic of lens positions in imaged mice. (D–L) N=10 (7M, 3 F) scope-mounted mice. (D) % time in dark. (E) Average speed. Wilcoxon L vs D, **p=0.0098. (F) Maximum speed. (G) Speed distribution normalized to zone. % time ≤2 cm/s (bar graph) Wilcoxon L vs D, *p=0.0137. (H) Transition speed. (I) Generalized Linear Mixed-Effect (GLME) fixed effects 95% CI (cm/s) in brackets. Approach speed is unchanged by body-in-light (BIL) [0.2506,–0.26603]. Retreat speed is increased by BIL [1.576, 0.9505]. (J) Distribution of zone and/or speed relationships among all recorded neurons. (K) Relationship between zone preference and speed encoding χ2 (3, n=532)=78.63, p<0.0001 (L) Distribution of zone and/or speed relationships among all transition-active neurons. (M) Distribution of zone and/or deceleration relationships among all transition-active neurons.
Figure 3
Light sensitivity and speed modulation are reflected in transition-active neurons.
Single-cell calcium imaging with miniscopes. n=10 (7M,3F) 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 body-in-light (BIL) or body-in-dark (BID). (D) Overlay of average ∆F/F (green, z-score) with mean transition speed (gray scale, cm/s). (E) Effect of BIL while approaching or retreating from zone transitions, GLME fixed effects 95% CI (a.u.). (F, G) 95% CI shown in red. (F) Mean R value per mouse for all acceleration events with significant (p<0.05) ∆F/F cross-correlation to speed (n=9 into light, 9 into dark, 10 within light, 10 within dark), or (G) acceleration (n=3 into light, 5 into dark, 8 within light, 9 within dark).
Figure 4 with 1 supplement
Patchy spiny projection neuron (SPN) efferents to substantia nigra pars reticulata (SNr) encode speed and deceleration.
Sepw1-Cre: n=8 (4M,4F) GPe, n=9 (5M,4F) GPi, n=8 (4M,4F) SNr; Calb1-Cre: n=8 SNr. 95% CI shown in red. (A) Schematic of AAV1-phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP injection into the dorsal striatum of a Sepw1-Cre mouse. A representative sagittal section shows patchy SPNs cell bodies and their efferents in the DS, GPe, GPi, and SNr from >3 mice. Scale bar 500 µm. (B) Injection schematic for mice with later fiber implants at GPe, GPi, or SNr. (C) Patchy SPN efferent activity aligned to zone transition, overlaid with speed. GCaMP8s into Light (light green) or Dark (dark green); simultaneous 405 nm channel into Light (light blue) or Dark (dark blue); mean speed (cm/s) during transition into Light (gray) or Dark (black). (D) Average across mice of mean R-value for acceleration events significantly correlated to ∆F/F (p<0.05) during zone transitions. (E–H) Patchy SPN ∆F/F at SNr cross-correlation to speed or acceleration aligned to acceleration events. Cross-correlogram color code: transition into Light (light green) or Dark (dark green), or within Light (light blue) or Dark (dark blue). (E) Heat map of cross-correlation to speed for each mouse. (F) ∆F/F cross-correlogram to speed. (G) Heat map of cross-correlation to acceleration for each mouse. (H) ∆F/F cross-correlogram to acceleration. (I–L) 95% CI in red. (I, J) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to speed, (I) mean R value per mouse, (J) mean Xcorr lag per mouse. (K, L) for all acceleration events with significant (p<0.05) ∆F/F cross-correlation to acceleration, (K) mean R value per mouse, (L) mean Xcorr lag per mouse. (M) Patchy SPN efferent ∆F/F at SNr inter-event interval in either zone, paired t-test, ***p=0.0001. (N–Q) Matrix ∆F/F at SNr (n=8; 4 M,4F). (N) Matrix efferent activity at SNr aligned to zone transition, overlaid with speed. Color coding identical to C. (O) Average across mice of mean R-value for acceleration events significantly correlated to ∆F/F (p<0.05) during zone transitions. (P, Q) Cross-correlogram color code identical to F and H. (P) ∆F/F cross-correlogram to speed. (Q) ∆F/F cross-correlogram to acceleration. (R) Matrix efferent ∆F/F at SNr inter-event interval in either zone, paired t-test, **p=0.0039.
Figure 4—figure supplement 1
Fiber photometry supplemental.
(A–K) N=8 (4M, 4 F) GPe-implanted, 9 (5M, 4 F) GPi-implanted Sepw1-Cre mice. (A) Representative images show GCaMP8s (green) and TH (magenta) immunostaining. N>3. Scale bar: 500 mm. (B) Transition body-in-light (BIL) effect on patchy spiny projection neuron (SPN) efferents. GLME fixed effects 95% CI (%∆F/F) in brackets. (C–F) Analysis of patchy SPN efferents in GPe. 95% CI in red. (C) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to speed, mean R value per mouse, (D) Mean Xcorr lag per mouse. (E) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to acceleration, mean R value per mouse. (F) Event frequent in either zone, paired t-test, **p=0.0018. (G–K) Analysis of patchy SPN efferents in GPi. 95% CI in red. (G) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to speed, mean R value per mouse. (H) Mean Xcorr lag per mouse. (I) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to acceleration, mean R value per mouse. (J) Mean Xcorr lag per mouse. (K) Event frequent in either zone, paired t-test, **p=0.0035. (L) N=9 (5M, 4 F) GRIN-implanted Sepw1-Cre mice. From net fluorescence collected in dorsal striatum, event frequent in either zone, paired, t-test *p=0.0373. (M–Q) N=8 (4M, 4 F) SNr-implanted Calb1-Cre mice. (M) Representative images show GCaMP8s (green) and TH (magenta) immunostaining. Scale bar: 500 mm. (N) Transition BIL effect on matrix efferents. Generalized Linear Mixed-Effect (GLME) fixed effects 95% CI (%∆F/F) in brackets. (O–Q) Analysis of matrix efferents in substantia nigra pars reticulata (SNr). 95% CI in red. (O) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to speed, mean R value per mouse. (P) Mean Xcorr lag per mouse. (Q) For all acceleration events with significant (p<0.05) ∆F/F cross-correlation to acceleration, mean R value per mouse.
Figure 5 with 1 supplement
Patchy spiny projection neuron (SPN) enhancement increases rest and eliminates discriminative speed at choice points.
(A) Injection schematic. (B) Dorsal striatal DREADD-mCherry overlaps patch marker MOR1 from >3 mice. Scale bar: 500 μm. 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–J) Light/Dark box. n=17 (9M,8F) Ctrl and 20 (12M,8F) 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. (G) Speed distribution normalized to zone. Insert, % time ≤2 cm/s 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–J) Transition speed for controls (H) and Gq (I). (J) 95% C.I. for Generalized Linear Mixed-Effect (GLME) by group shows speed approaching transitions is increased by body-in-light (BIL) for controls but reduced for Gq-mice. (K) Cartoon summarizing speed modulation by valence under control (blue) SA (red) and Gq (green) conditions.
Figure 5—figure supplement 1
Chemogenetics supplemental.
(A–E) Gi-mice L/D box, n=17 (9M, 8 F) Ctrl and 15 (8M, 7 F) Gi mice. (A) % time in the dark. Mann-Whitney, Ctrl vs Gi p=0.8232. (B) Average speed. Two-way RM ANOVA: group F (1, 30)=0.03006, p>0.05; light F (1, 30)=134.3, p<0.0001. (C) Maximum speed. Two-tailed Mann-Whitney, p=0.715. (D) Speed distribution normalized to zone. Insert for % time ≤2 cm/s (bar graph) paired t-test, L vs D Ctrl p<0.0001, Gi p<0.0001, unpaired t-test, Ctrl vs Gi in Light p>0.05, in Dark p>0.05. (E) Transition speed. Generalized Linear Mixed-Effect (GLME) fixed effects 95% CI (cm/s) in brackets. Approach speed is increased by body-in-light (BIL) [1.0773, 0.59214] and unchanged by Gi [–1.0473, 1.0196]; Retreat speed is increased by BIL [1.0562, 1.486] and unchanged by Gi [–1.3062, 0.92042]. GLME by group shows approach speed is increased by BIL for control [1.9076, 1.3185] and Gi [2.2463, 1.5553]. (F–I) Gi-mice LL/DD box. n=mouse (16 Ctrl, 13 Gi). (F) Average speed. Two-way RM ANOVA: lighting F (1, 27)=3,651 p=0.0667, group F (1, 27)=2.994 p>0.05. (G) Maximum speed. Two-way RM ANOVA: lighting F (1, 27)=0.05506 p>0.05, group F (1, 27)=1.402 p>0.05. (H) Speed distribution normalized to zone. For %time ≤2 cm/s (bar graph), Wilcoxon LL vs DD Ctrl p>0.05, Gi p=0.0105, Mann-Whitney, Ctrl vs Gi in Light p>0.05, in Dark p>0.05. (I) Transition speed. GLME fixed effects 95% CI (cm/s) in brackets. Approach speed is slightly increased by BIA [0.85304, 0.42385], unchanged by Gi [–0.25536, 1.985]; Retreat speed is slightly increased by BIA [0.31052, 0.76904], unchanged by Gi [–0.73445, 1.7233]. (J–M) Gq-mice LL/DD box. n=mouse (16 Ctrl, 19 Gq). (J) Average speed. Two-way RM ANOVA: lighting F (1, 33)=2.676 p>0.05, group F (1, 33)=1.343 p>0.05. (K) Maximum speed. Two-way RM ANOVA: lighting F (1, 33)=2.683 p>0.05, group F (1, 33)=0.0008 p>0.05. (L) Speed distribution normalized to zone. For %time ≤2 cm/s, Wilcoxon LL vs DD Ctrl p>0.05, Gq p>0.05, Mann-Whitney Ctrl vs Gq in LL p=0.0441, in DD p=0.0288. (M) Transition speed. GLME fixed effects 95% CI (cm/s) in brackets. Approach speed is increased by BIA [0.85304, 0.42385], unchanged by Gq [–1.1387, 0.89713]; Retreat speed is increased by BIA [0.31052, 0.76904], unchanged by Gq [–1.5338, 0.69961].
Figure 6
Patch/matrix balance at substantia nigra pars reticulata (SNr) controls Light/dark box locomotor phenotype.
(A) Injection schematic. (B) Sample image of TH (magenta) and mCherry (red) staining in SN from >3 mice. The fiber track is indicated with dashed line. Scale bars: 500 μm. (C–I) N=14 (7M,7F) Ctrl and 16 (8M,8F) 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. (F) Speed distribution normalized to zone. Insert, %time ≤2 cm/s L vs D Ctrl ***p=0.0002, ChR ***p=0.0005; Ctrl vs ChR in either zone p>0.05. (G–I) Transition speed for controls (G) and ChR (H). (I) 95% CI coefficient for Generalized Linear Mixed-Effect (GLME) by group shows speed approaching transitions is increased by body-in-light (BIL) for controls significantly more than for ChR mice. (J) Cartoon summarizing speed modulation by valence under control (blue) Gq (green) and SNr ChR (purple) conditions for SpCre mice. (K) injection schematic. (L) Sample image of TH (magenta) and mCherry (red) staining in SN. The fiber track is indicated with dashed line. Scale bars: 500 μm. (M–S) n=13 (7M,6F) Ctrl and 13 (7M,6F) ChR Calb1-Cre mice. (M) % time in dark, p>0.05. (N) 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. (O) Maximum speed. p>0.05. (P) Speed distribution normalized to zone. Insert, %time ≤2 cm/s L vs D Ctrl ****p<0.0001, ChR p>0.05; Ctrl vs ChR in either zone p>0.05. (Q–S) Transition speed for controls (Q) and ChR (R). (S) 95% CI coefficient for GLME by group. (T) Cartoon summarizing speed modulation by valence under control (blue) SA (red) and Calb1-Cre SNr ChR (maroon) conditions.
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Supplementary file 1
Statistics Table.
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Source data 1
Original data for each graph.
- https://cdn.elifesciences.org/articles/106403/elife-106403-data1-v1.xlsx
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Patchy striatonigral neurons modulate locomotor vigor in response to environmental valence
eLife 14:RP106403.
https://doi.org/10.7554/eLife.106403.4
