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. 2021 Dec 9;14:779436. doi: 10.3389/fnmol.2021.779436

FIGURE 1.

FIGURE 1

Experimental design. (A) Schematic diagram of the virus injection area. (B) Experimental design timeline. A total of 336 mice [including 144 D1R-cre mice and 24 wild-type littermate (WT) mice, 144 D2R-cre mice, and 24 WT mice] participated in this experiment. After adaptive feeding for 1 week, they were randomly divided into four large groups (84 mice in each large group), among which 72 D1R-cre mice and 12 WT mice were further divided into two small groups, and 72 D2R-cre mice and 12 WT littermate mice were further divided into two small groups. On the first day, one small group of D1R-cre mice and WT littermate mice, one small group of D2R-cre mice and WT mice were injected with the virus in the bilateral dSTR (200 nL/side), the other two small groups were injected with the virus in the bilateral SNpc (200 nL/side). Eighty-four mice in each large group were randomly divided into 14 groups according to the injections of different drugs or virus, including WT + Vehicle + mCherry + i.p. Saline (group a), D1R or D2R-cre + Vehicle + mCherry + i.p. Saline group (group b), D1R or D2R-cre + Vehicle + Gq + i.p. Saline group (group c), D1R or D2R-cre + Vehicle + Gi + i.p. Saline group (group d), D1R or D2R-cre + IDPN + mCherry + i.p. Saline group (group e), D1R or D2R-cre + IDPN + Gq + i.p. Saline group (group f), D1R or D2R-cre + IDPN + Gi + i.p. Saline group (group g), WT + Vehicle + mCherry + i.p. CNO group (group h), D1R or D2R-cre + Vehicle + mCherry + i.p. CNO group (group i), D1R or D2R-cre + Vehicle + Gq + i.p. CNO group (group j), D1R or D2R-cre + Vehicle + Gi + i.p. CNO group (group k), D1R or D2R-cre + IDPN + mCherry + i.p. CNO group (group l), D1R or D2R-cre + IDPN + Gq + i.p. CNO group (group m) and D1R or D2R-cre + IDPN + Gi + i.p. CNO group (group n) (n = 6 mice in each group). Mice in the groups a, b, e, h, i, and l were injected with a control virus (mCherry). Mice in the groups c, f, j, and m were injected with hM3Dq. Mice in group d, g, k, and n were injected with hM4Di. After the virus injection, all mice were back to the cage to rest for 4 days (days 2–5). On days 6–12, mice in groups e, f, g, l, m, and n were intraperitoneally injected with IDPN at 10:00 a.m. once daily, and mice in the other groups were intraperitoneally injected with 0.9% saline (Vehicle, contrast with IDPN). On the 23rd day, mice in the a–g groups were intraperitoneally injected with 0.9% saline (contrast with CNO), and mice in the h–n groups were intraperitoneally injected with clozapine N-oxide (CNO, Sigma, St. Louis, MO, United States, 1 mg/kg). Thirty minutes later, behavioral tests were carried out to compare the differences between the groups. Brain tissues for immunofluorescence labeled were collected on day 24.