Extended Data Fig. 1: PF projections to CPu, NAc, and STN, and PFNAc neurons have distinct electrophysiological properties as compared to PFCPu and PFSTN populations.
From: Targeting thalamic circuits rescues motor and mood deficits in PD mice
a-b, Anterograde tracing from mouse PF, ChR2-eYFP virus injection site (a), and PF terminals in CPu, NAc, and STN (b). DAPI staining (blue). c, CTB488 injection site in CPu (left) along with corresponding upstream PF labeling (lower magnification image relative to Fig. 1a, right). d, Upstream labeling in PF with 1 (CTB555) vs. 3 (CTB488) injections in CPu. 80.5% of CPu-projecting PF neurons (i.e., CTB488) are labeled by a single injection of CTB555 in CPu (n = 3 mice). e-f, Injection sites of CTB555 in STN (left) (e) and CTB633 in NAc (left) (f) along with corresponding upstream PF labeling images (lower magnification relative to Fig. 1a, both right panels). g-i, Representative high magnification images from Fig. 1a, showing low overlap between CPu- and NAc-projecting PF neurons (g), CPu- and STN-projecting PF neurons (h), and STN- and NAc-projecting PF neurons (i). j, Percentage of CTB488+ only (CPu only), CTB555+ only (STN only), and CTB633+ only (NAc only) in PF. Dashed lines indicate chance level (20.49% for CPu only, 15.7% for STN only, 8.28% for NAc only) calculated using NEUN staining (n = 4 mice per group). k, Percentage of NEUN+ PF neurons projecting to the three different targets (n = 4 mice per group). l-p, Electrophysiological properties of PFCPu, PFNAc, and PFSTN neurons, which were labeled using retrograde RV injected into CPu, NAc, and STN respectively. Input resistance (Rin) (l), membrane time constant (tau) (m), membrane capacitance (Cm) (n), action potential amplitude (o), and action potential afterhyperpolarization (AHP) amplitude (p) (For Rin, tau, and Cm, PFCPu: n = 20 neurons (7, 7, 6), PFSTN: n = 17 neurons (6, 5, 6), PFNAc: n = 19 neurons (6, 7, 6) from 3 mice each. For amplitude and AHP, PFCPu: n = 18 neurons (5, 5, 8), PFSTN: n = 16 neurons (6, 5, 5), PFNAc: n = 19 neurons (6, 7, 6) from 3 mice each). q, ChR2-eYFP virus was injected in PF and ex vivo recordings were performed at PF cell bodies to validate virus expression (left), light responses in ChR2-labeled PF neurons were evoked by a 473 nm pulse train, shown in both current and voltage clamp modes (right). r, Reliability of ChR2 was compared across PF populations by recording from retrogradely-labeled CTB+ neurons that also expressed ChR2 (n = 12 neurons (4, 4, 4) from 3 mice for each group). s, Onset latency of evoked EPSCs recorded in postsynaptic neurons receiving PF input (PF→CPu: n = 19 neurons (7, 6, 6), PF→STN: n = 26 neurons (10, 9, 7), PF→NAc: n = 20 neurons (7, 7, 6) from 3 mice each). Data are presented as mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant. One-sample t test (j), one-way ANOVA followed by Bonferroni post-hoc test (k-p, s), and two-way ANOVA with repeated measures followed by Bonferroni post-hoc test (r). CPu only P = 0.033, STN only P = 0.028, NAc only P = 0.006 (j), F = 8.88, P = 0.007, CPu vs. STN t = 1.4, STN vs. NAc t = 2.74, CPu vs. NAc t = 4.14 (k), F = 27.71, P < 0.0001, PFCPu vs. PFNAc t = 6.57, PFSTN vs. PFNAc t = 6.28 (l), F = 17.48, P < 0.0001, PFCPu vs. PFNAc t = 5.17, PFSTN vs. PFNAc t = 5.04 (m), F = 11.76, P < 0.0001, PFCPu vs. PFNAc t = 3.37, PFSTN vs. PFNAc t = 4.69 (n), F = 7.19, P = 0.0018, PFCPu vs. PFNAc t = 3.78 (o), F = 6.11, P = 0.0042, PFCPu vs. PFNAc t = 3.26, PFSTN vs. PFNAc t = 2.65 (p), Amplitude: F = 0.03, DFn = 2, DFd = 297, P = 0.97, spikes/stimulus: F = 0.68, DFn = 2, DFd = 297, P = 0.51 (r), F = 0.11, P = 0.89 (s)
