Extended Data Fig. 2: PF circuit electrophysiology, PF→STN terminal inhibition during motor learning, and cell type-specific tracing for the PF→CPu circuit.
From: Targeting thalamic circuits rescues motor and mood deficits in PD mice
a-c, Representative traces and quantification of evoked EPSCs in the presence of tetrodotoxin (TTX), 4-aminopyridine (4AP), and 6-cyano-7-nitroquinovaline-2,3-dione (CNQX) for PF→CPu (a), PF→STN (b), and PF→NAc (c) circuits (PF→CPu: n = 4 neurons (2, 1, 1), PF→STN: n = 3 neurons (1, 1, 1), PF→NAc: n = 3 neurons (1, 1, 1) from 3 mice each). Norm. peak EPSC amplitude plotted. d, Representative traces and quantification of paired-pulse ratio (also referred to as short-term plasticity) recordings in PF circuits (PF→CPu: n = 20 neurons (7, 6, 7), PF→STN: n = 23 neurons (8, 7, 8), PF→NAc: n = 20 neurons (7, 6, 7) from 3 mice each). e, C21-induced, reversible neuronal inhibition of a PF neuron expressing hM4Di ex vivo, using a step current injection protocol. f, C21-induced inhibition of PFCPu or PFSTN neurons during rotarod tests (n = 14 mCh, n = 19 PFCPu, n = 18 PFSTN mice). g, Experimental protocol for AMPA/NMDA ratio recordings. CaMKII-ChR2-eYFP virus was injected in PF, 3 weeks later animals were trained on the rotarod paradigm, and 1 hr after the end of training ex vivo recordings were performed. h, CaMKII-eArch3.0-eYFP virus was injected in PF and fibers were implanted above STN. eArch-eYFP virus expression in PF (left), and light-induced neuronal inhibition ex vivo (right). i, PF→STN terminal inhibition followed by cFos staining in STN using home cage or rotarod mice validated effective in vivo terminal inhibition (n = 4 mice per group). cFos was stained using a 633 secondary antibody (pseudocolored red). j, PF→STN terminal inhibition during rotarod. eYFP control mice received a CaMKII-eYFP virus in PF in place of the eArch virus. Norm. latency plotted relative to day 1 (n = 10 eYFP, n = 9 eArch mice). k-m, Monosynaptic retrograde RV tracing from D1+ or D2+ MSNs in CPu. Images show starter cells (k), FISH co-staining of GFP with D1 or D2 (l), and corresponding PF labeling (m) (n = 6 D1-Cre, n = 5 D2-Cre mice). n, Representative traces (left) and current-frequency curves (right) of ex vivo recordings from D2- (putative D1+) or D2+ MSNs in CPu (D1: n = 14 neurons (5, 4, 5), D2: n = 12 neurons (4, 5, 3) from 3 mice each). o, Anterior to posterior (AP) distribution of PV neurons in STN. Cre-dependent mCh virus was injected into STN of PV-Cre mice. Data are presented as mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant. One-way ANOVA followed by Bonferroni post-hoc test (a-c, f), two-way ANOVA with repeated measures followed by Bonferroni post-hoc test (d, n), unpaired t test (i, m), and two-tailed paired t test (j). F = 24.64, P < 0.0001, Baseline vs. TTX t = 4.56, TTX vs. TTX + 4AP t = 7.23, TTX + 4AP vs. TTX + 4AP + CNQX t = 6.99 (a), F = 9.59, P = 0.005, TTX vs. TTX + 4AP t = 4.71, TTX + 4AP vs. TTX + 4AP + CNQX t = 4.57 (b), F = 468.7, P < 0.0001, Baseline vs. TTX t = 22.74, TTX vs. TTX + 4AP t = 30.83, TTX + 4AP vs. TTX + 4AP + CNQX t = 28.97 (c), F = 6.41, DFn = 2, DFd = 120, P = 0.003 (d), Day 1: F = 11.30, P < 0.0001, mCh vs. PFCPu t = 3.25, mCh vs. PFSTN t = 1.00, Day 2: F = 22.58, P < 0.0001, mCh vs. PFCPu t = 1.70, mCh vs. PFSTN t = 4.38 (f), mCh P = 0.012, eArch P = 0.55 (i), eYFP P = 0.0032, eArch P = 0.24 (j), P = 0.61 (m), F = 28.31, DFn = 1, DFd = 240, P < 0.0001 (n)
