Extended Data Fig. 9: Testing molecular target candidates in the PF→STN and PF→CPu circuits.

a, GUCD1 (guanylyl cyclase gene, the NO receptor), ERBB4, and OPRM1 (μ-opioid receptor gene) FISH staining with PV in STN sections. 65.6% of all PV⁺ neurons were GUCD1⁺, 58.5% of all PV⁺ neurons were ERBB4⁺, 57.2% of all PV⁺ neurons were OPRM1⁺(n = 3 mice per group). b, AMPA/NMDA ratio recordings of the PF→STN circuit, specifically PV⁺ STN neurons, before and after optical high frequency stimulation (HFS) in PD model mice (n = 8 neurons (2, 3, 3) from 3 mice). oChIEF-mCh virus was injected in PF, Cre-dependent eYFP virus in STN, and 6-OHDA in SNc of PV-Cre mice. c-d, With DAMGO (μ-opioid receptor agonist), SNP (NO receptor agonist), AG1478 (ERBB4 receptor antagonist) (c) or PNU282987 (α7 agonist) (d) bath application, AMPA/NMDA ratio recordings of the PF→STN_PV circuit before and after optical HFS in PD model mice (DAMGO: n = 7 neurons (2, 2, 3), SNP: n = 7 neurons (3, 2, 2), AG1478: n = 7 neurons (2, 2, 3), PNU282987: n = 8 neurons (2, 3, 3) from 3 mice each). oChIEF-mCh virus was injected in PF, Cre-dependent eYFP virus in STN, and 6-OHDA in SNc of PV-Cre mice. e, Local bilateral infusion of PNU282987, α-Ctx MII, or epibatidine into STN of PD model mice prior to rotarod behavior assays (n = 8 mice per group). f, α7 nAChRs FISH staining with PV in STN sections from mCh and KD mice, α7 in PV⁺ cells is decreased by 84% in KD mice as compared to mCh controls (n = 3 mCh, n = 4 α7 KD mice). g, Activating α7 nAChRs without (left) or with (right) α7 KD from PV⁺ STN neurons during rotarod, using PD model mice (n = 10 mCh, n = 8 α7 KD mice). PD_{α7 KD} received a Cre-dependent Cas9 and gRNA viruses (for knockdown of α7 nAChRs) and cannula implants above STN, and 6-OHDA in SNc of PV-Cre mice. h, α6 nAChRs FISH staining with D1 or D2 in mouse CPu sections and with PF_CPu neurons in mouse PF sections. PF_CPu neurons were labeled by injecting AAVretro-GFP in CPu (n = 3 D1⁺, n = 3 D2⁺, n = 4 PF_CPu⁺ mice). i, Representative cannula implant targeting CPu for local infusion experiments (left), local bilateral infusion of PBS, PNU282987, or epibatidine into CPu of PD model mice and infusion of PBS into CPu of WT mice prior to open field behavior (n = 10 WT_PBS, 9 PD_PBS, 9 PD_PNU, 9 PD_Epibat mice) (right). WT_PBS and PD_PBS data from Fig. 5e. j, α6 nAChRs FISH staining with PF_CPu neurons in PF sections from mCh and KD mice, α6 in PF_CPu⁺ cells is decreased by 78% in KD mice as compared to mCh controls (n = 4 mice per group). k, Total distance mice travelled in the open field after KD of α6 in PF_CPu neurons of PD model mice (n = 8 mice per group). PD_{α6 KD} received a retrograde RVdGL-Cre in CPu, Cre-dependent Cas9 and gRNA viruses (for knockdown of α6 nAChRs) in PF_CPu neurons, and 6-OHDA in SNc. Data are presented as mean ± SEM; *P < 0.05, ***P < 0.001. NS, not significant. One-way ANOVA followed by Bonferroni post-hoc test (a, h, i), two-tailed paired t test (b-e, g), and two-tailed unpaired t test (f, j, k). F = 0.45, P = 0.13 (a), P = 0.60 (b), DAMGO P = 0.48, SNP P = 0.82, AG1478 P = 0.12 (c), P = 0.011 (d), PNU282987 P = 0.034, α-Ctx MII P = 0.34, Epibatidine P = 0.66 (e), P < 0.0001 (f), mCh + PNU282987 P = 0.04, α7 KD + PNU282987 P = 0.37 (g), F = 456.9, P < 0.0001, D1⁺ vs. PF_CPu⁺ t = 25.49, D2⁺ vs. PF_CPu⁺ t = 25.6 (h), F = 5.16, P = 0.0049, WT_PBS vs. PD_PBS t = 3.45, WT_PBS vs. PD_PNU t = 3.09, WT_PBS vs. PD_Epibat t = 2.86 (i), P < 0.0001 (j), P = 0.02 (k)

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