Figures and data
dSPNs exhibit spatially clustered activity that varies with movement.
(a) Cartoon representation of implanted GRIN lens and miniscope imaging in the DLS. (b) Examples of segmented cell outlines from the CNMF-E algorithm in a single mouse. (c) Example of Ca2+ traces from corresponding colored and filled segmented cells shown in (b). (d) The probability of a Ca2+ event in detected cells increases during movement in dSPNs. (e) The Ca2+ event rate in detected cells increases during movement in dSPNs (p=0.240, Wilcoxon Signed Rank Test, events/second during bins with velocity>0.5 cm/sec vs. during bins with velocity<0.5 cm/sec). (f) Cartoon schematic to illustrate analysis of co-activity as a function of distance between cell pairs. Across all cell pairs in the field-of-view for each session, a correlation matrix was generated, in which, for each frame of the recording, the cell pair was scored a 1 if both cells were active, or 0 otherwise. These scores for each frame were averaged across all frames in the recording, to generate an average co-activity score for that pair. Then, those values were normalized to averages measured from independently shuffled Ca2+ traces for each cell in the recording (using 1000 independent shuffles for each cell trace), to control for changes in mean activity level between conditions. (g) Hypothetical plot of shuffle-normalized jaccard scores versus distance when coactivity is not “clustered” and hypothetical plot of “clustered” co-activity. Co-activity decreases during periods of movement (Linear Mixed Effects Model/LMM, p<0.000, z=-4.852, β=-1.671, SE = 0.344, 95% CI: [−2.346 −0.996], N=5 mice). (i) The spatial coordination index (SCI) peaks during movement onset (p<0.000, z=12.21, Wilcoxon Signed Rank Test, comparing the peak value of the normalized SCI during 2.5 seconds following movement onset to 1).
A negative allosteric modulator of mGluR5 increases coactivity among dSPN pairs.
(a) Cartoon representing implanted GRIN lens and miniscope imaging in the DLS (b) Examples of segmented cell outlines from the CNMF-E algorithm in a single mouse. (c) Example of Ca2+ traces from corresponding filled segmented cells shown in (b) (d) Binned velocity of animal. Fenobam increased the amount of time the animal spends at rest (Vehicle mean: 0.324 ± 0.091, fenobam mean: 0.635 ± 0.043, p=0.024, U=10, Mann-Whitney U test) (e) Mean velocity of animals following vehicle or fenobam treatment. The fenobam effect on mean velocity during the open field test is not significant (Mann-Whitney U test, p=0.103, U=48) (f) Mean bout length of animals during vehicle or fenobam. Fenobam reduces the mean bout length (Vehicle: 7.098 seconds ± 1.272, Fenobam: 2.886 seconds ± 0.160, Mann-Whitney U test, p<0.001, U=0), but does not affect the number of bouts (Mann-Whitney U test, p=0.793, U=35). (g) The average event rate of dSPNs at each movement velocity of the mouse. This relationship is not affected by fenobam administration (Linear Mixed Effects Model/LMM, p=0.754, z=-0.313, β=-0.002, SE = 0.006, 95% CI: [−0.014 0.010], N=8 mice). (h) Fenobam increases the pairwise coactivity of dSPNs when grouped into 50 μm bins (LMM, p<0.000, z=-6.050, β=-0.828, SE = 0.137, 95% CI: [−1.096 −0.560], N=8 mice) (i) Following both vehicle and drug treatment spatial coordination peaks during movement onset (Veh: p<0.000, z=12.21, Drug: p<0.000, z=10.1804) (j) Pairwise coactivity during rest with vehicle and fenobam treatment (velocity <0.5 cm*sec-1)(LMM, p<0.000, z=-4.448, β=-1.292, SE = 0.290, 95% CI: [−1.861 −0.723], N=8 mice) (k) Pairwise coactivity during movement (velocity >0.5 cm*sec-1) with vehicle and fenobam (LMM, p=0.008, z=-2.673, β=-0.220, SE = 0.082, 95% CI: [−0.381 −0.059], N=8 mice) (l) Fenobam does not affect the relative decrease in coactivity seen during movement
A positive allosteric modulator of mGluR5, JNJ-46778212, decreases clustered coactivity of dSPNs.
(a) Time animal spent in each velocity bin and (b) Mean velocity of the animals during vehicle and after JNJ administration. JNJ decreases the amount of time the animal spends at rest (Vehicle mean: 0.661 ± 0.066, JNJ mean: 0.294 ± 0.041, p=0.012, U=25, Mann-Whitney U test), however the change in mean velocity (b) is not significant (p=0.060, U=3, Mann-Whitney U test), nor is the mean bout length during vehicle and JNJ (p=0.060, U=3, Mann-Whitney U test) (d) Comparison of mean number of bouts of each animal during vehicle and JNJ (p=0.403, U=8, Mann-Whitney U test). (e) Mean event rate of Ca2+ transients in dSPNs during movement. JNJ administration does not significantly affect the event rate in dSPNs (LMM, p=0.058, z=1.893, β=0.010, SE = 0.005, 95% CI: [−0.000 0.020], N=4 mice) (f) Coactivity of pairs of dSPNs (50 μm bins) during total time in open field. Systemic JNJ administration reduces pairwise coactivity (LMM, p=0.003, z=2.964, β=-0.450, SE = 0.152, 95% CI: [−0.153 0.748], N=4 mice) (g) Spatial coordination increases at movement onset in both vehicle and drug groups (VEH: p<0.000, z=6.249, JNJ: p<0.000, z=9.111, Wilcoxon Signed Rank test) (i) Normalized co-activity of dSPNs during rest. JNJ produces a small but significant reduction in coactivity measured during rest (velocity < 0.5 cm*sec-1)(LMM, p=0.030, z=2.165, β=0.618, SE = 0.285, 95% CI: [0.059 1.177], N=4 mice) (j) Normalized co-activity of dSPNs during movement (velocity > 0.5 cm*sec-1). JNJ had a significant effect on dSPN clustered activity (LMM, p=0.027, z=-2.212, β=-1.694, SE = 0.766, 95% CI: [−3.195 −0.193], N=4 mice).(k) Event rate in dSPNs during vehicle and after JZL administration. JZL-184 administration reduces the event rate (LMM, p=0.001, z=3.195, β=0.007, SE = 0.002, 95% CI: [0.003 0.012], N=5 mice) (l) Normalized co-activity of dSPNs against Euclidean distance. JZL had no effect on coactivity (LMM, p=0.065, z=-1.847, β=-1.204, SE = 0.652, 95% CI: [−2.481 0.074], N=5 mice) (m) Time animals spent in velocity bins. JZL did not affect time spent at rest (p=0.531, U=16, Mann-Whitney U test).
mGluR5 cKO in Drd1a-expressing neurons reduces spontaneous motor behaviors and affects synaptic properties of dSPNs.
(a) (Top) Cartoon representation of the role of mGluR5 in mobilizing eCBs at corticostriatal synapses. (Bottom) Representative EPSCs recorded prior to and during DHPG in WT mice (b) Example time course of corticostriatal EPSCs recorded in dSPNs during application of 100 μM DHPG in slice from WT mice. (c) Grouped data of DHPG depression. D1 cKO mice lack DHPG induced synaptic depression (cKO normalized EPSC amplitude: 0.754 ± 0.082, Wilcoxon Signed Rank Test, p=0.018, W=2; WT normalized amplitude: 0.992 ± 0.992, Wilcoxon Signed Rank Test p=0.834, W=9). (d) Time course of activity in open field and (e) Total distance traveled by WT and cKO mice. D1 cKO mice have reduced activity in the open field, measured as distance traveled over time and total distance traveled during a 30-minute open-field session (Mann-Whitney U Test, U=245, p<0.001, N = 16 WT, 17 cKO). (f) Digging time in WT and cKO mice. D1 cKO mice spend less time digging when placed into a novel home cage (Mann-Whitney U Test, U=51.5, p=0.008, N = 7 WT, 8 cKO). (g) Latency to fall in the accelerating rotarod test. dSPN cKO mice have consistently reduced latency during the accelerating rotarod test over 3 days, with 4 sessions per day (p=0.024, F=5.709, Two-way repeated measures ANOVA, N = 12 WT, 17 cKO). (h) Running distance per hour on wireless wheels placed in home cage over 7 days. (i) Total distance run per day during first 3 days and over 7 days. D1 cKO mice ran significantly less during the first 3 days of testing (Mann-Whitney U Test, U=173, p=0.049) (j) Example traces of voltage response to 200pA current injection in WT and cKO mice. (k) There is not significant difference in the input output curve for current injection and AP firing between genotypes (LMM, p=0.056, z=1.913, β=1.197, SE = 0.626, 95% CI: [−0.029 2.424], N=26 cells, WT, N=22 cells, cKO). (l) Example mEPSC recordings in dSPN from WT and D1 cKO mice (m) Average amplitude of mEPSCs and cumulative distribution of amplitudes in WT and D1 cKO mice. There is no difference in amplitudes in recordings from the two genotypes (WT AMP: 21.04 ± 1.71 pA, N=15, cKO AMP: 16.66 ± 1.12, N=8, Mann-Whitney U Test, U=86.5, p=0.093) (n) Average mEPSC frequency and cumulative interevent intervals in WT and D1 cKO animals. D1 cKO mice have increased mEPSC frequency (WT Hz: 1.920 ± 0.3319 pA, N=15, cKO Hz: 3.325 ± 0.525, N=8, Mann-Whitney U Test, U=27.5, p=0.039)
D1 cKO mice show cell autonomous increase in clustered coactivity in dSPNs.
(a) Cartoon representation of miniscope imaging in the DLS (b) Example segmented image from cKO mice (c) Example Ca2+ activity traces from identified dSPNs in cKO mice, filled cells in (b) (d) Mean event rate during movement in WT and cKO animals. The event rate in dSPN cKO mice is unchanged from WT (LMM, p=0.651, z=0.452, β=0.003, SE = 0.007, 95% CI: [−0.011 0.017], N=9 wt, 6 cKO) (e) Coactivity of pairs of dSPNs (50 μm bins) in WT and cKO animals. D1 cKO mice have increased coactivity across all pairwise distances (MLM, p=0.021, z=2.305, β=3.989, SE = 1.731, 95% CI: [0.597 7.382], N=9 wt, 6 cKO) (f) Spatial coordination peaks at movement onset in both wt and cko mice (WT: p<0.000, z = 7.301, KO: p<0.000, z = 6.700, Wilcoxon Signed Rank test) (g) The relative increase in coactivity during rest is not statistically different between cKO and wt mice (LMM, p=0.552, z=0.595, β=1.391, SE=2.339, 95% CI: [−3.193 5.976]) (h) iSPN event rate in D1 cKO and WT animals is unchanged (LMM, p=0.310, z=1.016, β=0.017, SE = 0.016, 95% CI: [−0.015 0.048], N=3 wt, 3 cKO) (i) Clustered coactivity of iSPNs in WT and D1 mGluR5 cKO animals is unaffected (LMM, p=0.691, z=0.398, β=2.368, SE = 5.953, 95% CI: [−9.300 14.035], N=3 wt, 3 cKO).
a) Averaged Ca2+ transients from vehicle and fenobam treated mice b) Fenobam treatment does not affect the distribution of event amplitudes (LMM, p=0.246, z=-1.160, β=-0.555, SE=0.478, 95% CI: [−1.493 0.383], N=8 mice) c) Fenobam treatment does not affect the distribution of event lengths (LMM, p=0.499, z=-0.676, β=-0.044, SE=0.065, 95% CI: [−0.173 0.084], N=8 mice).
a) Averaged Ca2+ transients from vehicle and JNJ treated mice b) JNJ treatment does not affect the distribution of event amplitudes (LMM, p=0.955, z=0.056, β=0.048, SE=0.863, 95% CI: [−1.644 1.741], N=5 mice) c) JNJ treatment does not affect the distribution of event lengths (LMM, p=0.913, z=0.110, β=0.021, SE=0.188, 95% CI: [−0.348 0.389], N=5 mice)
a) Averaged Ca2+ transients from wt and cKO mice b) Wt and cKO mice show similar distributions of event amplitudes (LMM, p=0.497, z=-0.679, β=-0.177, SE=0.261, 95% CI: [−0.690 0.335], N=5 mice) c) Wt and cKO mice show similar distributions of event length (LMM, p=0.284, z=1.072, β=0.140, SE=0.130, 95% CI: [−0.116 0.395], N=5 mice)
Number of cells per animal and recording sessions from animals used in figure 1 analysis.
Number of cells per animal and recording sessions from animals used in figure 2 analysis
Vehicle Sessions for JNJ experiments. Number of cells per animal and recording sessions from animals used in figure 3 analysis
