Activation of zona incerta gamma-aminobutyric acid-ergic neurons alleviates depression-like and anxiety-like behaviors induced by chronic restraint stress

Figure 3 Effects of chemogenetic manipulation of gamma-aminobutyric acid-ergic neurons in the zona incerta on chronic resistant stress mice behavior. A: Experimental strategy and timeline for behavioral assays; B: Bath application of clozapine-N-oxide (10 μM) induced neuronal discharge in neurons expressing hM3Dq-mCherry (clamped at -70 mV). It suppressed spiking in the hM4Di-mCherry group (clamped at -40 mV); C: The injection site of the virus in the zona incerta. Scale bars: 200 µm; D: Representative image of colocalization of neurons expressing mCherry and gamma-aminobutyric acid in the zona incerta. Blue: DAPI staining. Scale bars: 100 μm; E: Impact of chemogenetic manipulation on the time spent in the center of the open field test by mice in the chronic resistant stress (CRS) group; F: Impact of chemogenetic manipulation on the total distance traveled by mice in the CRS group in the open field test; G: Impact of chemogenetic manipulation on the time spent in the open arms of the elevated plus maze by mice in the CRS group; H: Impact of chemogenetic manipulation on the total distance traveled by mice in the CRS group in the elevated plus maze; I: Effect of chemogenetic manipulation on immobility time in the forced swim test for mice in the CRS group; J: Effect of chemogenetic manipulation on immobility time in the tail suspension test for mice in the CRS group; K and L: Effect of chemogenetic manipulation on sucrose preference and total liquid consumption in the sucrose preference test for mice in the CRS group. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001; n = 12 for the EGFP group and hM3Dq group; n = 11 for hM4Di group; 3v: 3rd ventricle; NS: Not significant; CNO: Clozapine-N-oxide; GABA: Gamma-aminobutyric acid; OFT: Open field test; EPM: Elevated plus maze; FST: Forced swimming test; TST: Tail suspension test; SPT: Sucrose preference test.