Extended Data Fig. 10: Testing molecular target candidates in the PF→NAc circuit, and rescue of motor phenotypes in the same mice using PFCPu cell body and PF→STN circuit manipulations in vivo.
From: Targeting thalamic circuits rescues motor and mood deficits in PD mice
a, Before and after bath application of PNU282987 (α7 agonist), CC4 (α6 agonist), and UB165 (α3 agonist), evoked EPSCs were recorded from D1 MSNs in NAc of PD model mice. oChIEF-mCh virus was injected in PF and 6-OHDA in SNc of D2-eGFP mice (PNU282987: n = 6 neurons (1, 2, 1, 1, 1), CC4: n = 7 neurons (1, 2, 1, 1, 2), UB165: n = 8 neurons (2, 2, 1, 2, 1) from 5 mice each). b, Representative cannula implant targeting NAc for local infusion experiments (left), local bilateral infusion of PBS, α-Ctx MII, or PNU282987 into NAc of PD model mice and infusion of PBS into NAc of WT mice prior to depression-like behavior tests (n = 9 mice per group) (right). WTPBS and PDPBS data from Fig. 5h. c, β2 nAChRs FISH staining with D1 in NAc sections from mCh and KD mice, β2 in D1+ cells is decreased by 80% in KD mice as compared to mCh controls (n = 3 mCh, n = 4 β2 KD mice). d, Activating β2 nAChRs without (PDmCh+Epibat) or with (PDβ2 KD+Epibat) β2 KD from D1+ NAc neurons during depression-like behaviors, using PD model mice (n = 9 mice per group). Dashed line indicates WT level from Fig. 5h. PDβ2 KD received a Cre-dependent Cas9 and gRNA viruses (for knockdown of β2 nAChRs), cannula implants in NAc, and 6-OHDA in SNc of D1-Cre mice. e, A retrograde RVdGL-Cre was injected in CPu, Cre-dependent hM4Di-mCitrine (for PFCPu labeling), and CaMKII-oChIEF-mCh (for STN terminal manipulations) in PF, optic fibers targeting STN, and 6-OHDA injected in SNc. f, Total distance in the open field was measured 40 min after C21 injections. PD model mice that received 6-OHDA injections in SNc without any subsequent PF manipulation was the baseline group (labeled PD). PD model mice with PFCPu manipulations are referred to as PD Rescue (n = 11 PD, n = 8 PD Rescue mice). g, Rotarod tests were performed after the open field paradigm. For rotarod, PF→STN circuit strengthening using our optical LTP protocol (20 + 100 Hz) was performed (n = 11 PD, n = 8 PD Rescue mice). Data are presented as mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant. Two-tailed paired t test (a, g), one-way ANOVA followed by Bonferroni post-hoc test (b), and two-tailed unpaired t test (c, d, f). PNU282987 P = 0.14, CC4 P = 0.17, UB165 P = 0.10 (a), Sucrose preference: F = 8.71, P = 0.0002, WTPBS vs. PDPBS t = 4.57, WTPBS vs. PDα-Ctx MII t = 4.14, WTPBS vs. PDPNU282987 t = 3.57, Forced swim: F = 5.59, P = 0.0034, WTPBS vs. PDPBS t = 3.16, WTPBS vs. PDα-Ctx MII t = 3.32, WTPBS vs. PDPNU282987 t = 3.52, Tail suspension: F = 5.32, P = 0.0044, WTPBS vs. PDPBS t = 3.44, WTPBS vs. PDα-Ctx MII t = 3.28, WTPBS vs. PDPNU282987 t = 2.99 (b), P < 0.0001 (c), Sucrose preference P = 0.022, Forced swim P = 0.012, Tail suspension P = 0.029 (d), P = 0.0017 (f), PD P = 0.78, PD Rescue P = 0.011 (g)
