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. 2023 Sep 6;43(36):6249–6267. doi: 10.1523/JNEUROSCI.0247-23.2023

Figure 10.

Figure 10.

Attenuated hyperdopaminergic behaviors in DAT-Cul3f/+ mice by chemogenetic inhibition of midbrain DA neurons. A, Schematic diagram represents injection of AAV5-SunDIO-hM4D(Gi)-mCherry virus (hM4Di) into VTA in DAT-Cre or DAT-Cul3f/+ mouse (left) and experimental approach for Cre-dependent expression of hM4Di-mCherry (right). AAV5-DIO-mCherry (mCherry) virus was injected as control. B, Decreased distance traveled by hM4Di-injected DAT-Cul3f/+ mice in the open field test after CNO injection (2 mg/kg, i.p.). n = 12 mice for all groups; for DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.0177; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.0302; unpaired t test. C, Comparable time spent in the center in the open field after CNO injection. n = 12 mice for all groups; for DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.509; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.7284; unpaired t test. D, Increased spontaneous alternation in hM4Di-injected DAT-Cul3f/+ mice in Y-maze test after CNO injection. n = 12 mice for all groups; for DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.0015; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.0219; unpaired t test. E, Comparable total number of entries in Y-maze after CNO injection. n = 12 mice for all groups; for DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.5886; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.6722; unpaired t test. F, Restored PPI in hM4Di-injected DAT-Cul3f/+ mice after CNO injection. n = 12 mice for all groups. For 73 dB, DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.5555; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.0708. For 76 dB, DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.03; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.0310. For 82 dB, DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.0013; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.0342; unpaired t test. G, No impact on acoustic startle response by CNO for indicated groups. n = 12 mice for all groups; for DAT-Cre + hM4Di + veh versus DAT-Cul3f/+ + hM4Di + veh, p = 0.4624; DAT-Cul3f/+ + mCherry + CNO versus DAT-Cul3f/+ + hM4Di + CNO, p = 0.7720; unpaired t test. H, Post hoc staining to confirm the expression of the injected AAV5-DIO-hM4Di-mCherry virus. Top, Representative images of Cre-dependent expression of hM4Di-mCherry in DAT-Cul3f/+ mouse in the midbrain. The coronal section of the midbrain was stained with mCherry antibody (visualized by AlexaFluor-594, red) and DAPI (blue). Scale bar, 500 μm. Bottom, Higher magnification of the insets in top panels. Scale bar, 200 μm. IPN, interpeduncular nucleus. Data are mean ± SEM. *p < 0.05. **p < 0.01.