Selective knock-out of PI4KⅢβ in the VTA DA neurons reduces phosphoinositide levels and increases the susceptibility of the mice to stress stimulation. A, PI4KⅢβ^fl/fl mice were mated with DAT-Cre mice to establish a mouse line with PI4KⅢβ being selectively deleted in the DA neurons, the DAT^{PI4KⅢβ−/−} mice line. B, Single-cell PCR of the VTA DA neurons in the WT and the DAT^{PI4KⅢβ−/−} mice. C, Immunofluorescence of the WT and the DAT^{PI4KⅢβ−/−} mice, PI4KⅢβ labeling is in green, and TH labeling is in red. D, Western blot results of PI4KⅢβ in the VTA. N = 9 for the WT and the DAT^{PI4KⅢβ−/−} mice (t₍₁₆₎ = 4.362, ***p = 0.0005, unpaired t test). E, Normalized PIP₂ (i) and PIP (i) quantity (in ng) based on the quantity of total protein level (mg) for the WT and the DAT^{PI4KⅢβ−/−} mice. N = 12 for the WT mice and N = 9 for the DAT^{PI4KⅢβ−/−} mice (i, t₍₁₉₎ = 2.365, *p = 0.0288, unpaired t test; ii, t₍₁₉₎ = 2.689, *p = 0.0145, unpaired t test). F, Proportion of susceptible or resilient mice responding to SDS stimulation (i), N = 22 for the WT mice and N = 25 for the DAT^{PI4KⅢβ−/−} mice (<mml:math display="inline"><mml:msup><mml:mi>χ</mml:mi><mml:mn>2</mml:mn></mml:msup><mml:mo>=</mml:mo><mml:mn>5.144</mml:mn></mml:math>χ2=5.144, *p = 0.0233, chi-square test); SIR (ii), two-way (genotype, stress) repeated measures ANOVA: genotype, F_(1,45) = 0.3607, p = 0.5477; stress, F_(1,45) = 286.6, p < 0.0001. Bonferroni’s post hoc test: significant difference between pre-SDS and post-SDS, ****p < 0.0001; social interaction time (iii); sucrose consumption rate (iv). Two-way (genotype, stress) repeated measures ANOVA: genotype, F_(1,45) = 3.318, p = 0.0752; stress, F_(1,45) = 45.08, p < 0.0001. Bonferroni’s post hoc test: significant difference between pre-SDS and post-SDS, **p = 0.0037, ****p < 0.0001; significant difference between WT and DAT^{PI4KⅢβ−/−} mice, *p = 0.0259; N = 22 for the WT mice and N = 25 for the DAT^{PI4KⅢβ−/−} mice. G, Proportion of susceptible or resilient mice responding to a subthreshold SDS stimulation (i), N = 12 for the WT mice and N = 14 for the DAT^{PI4KⅢβ−/−} mice (<mml:math display="inline"><mml:msup><mml:mi>χ</mml:mi><mml:mn>2</mml:mn></mml:msup><mml:mo>=</mml:mo><mml:mn>4.013</mml:mn></mml:math>χ2=4.013, *p = 0.0452, chi-square test); SIR (ii). Two-way (genotype, stress) repeated measures ANOVA: genotype, F_(1,24) = 7.380, p = 0.0120; stress, F_(1,24) = 2.780, p = 0.1085. Bonferroni’s post hoc test: significant difference between pre-SDS and post-SDS, ***p = 0.0007, a significant difference between WT and DAT^{PI4KⅢβ−/−} mice, ****p < 0.0001; social interaction time (iii); sucrose consumption rate. Two-way (genotype, stress) repeated measures ANOVA: genotype, F_(1,24) = 4.672, p = 0.0408; stress, F_(1,24) = 0.5928, p = 0.4489. N = 12 for the WT mice and N = 14 for the DAT^{PI4KⅢβ−/−} mice. H, OFT of male mice. (i) The total distance traveled, N = 20 for the WT mice and N = 22 for the DAT^{PI4KIIIβ−/−} mice (t_(31.44) = 3.036, **p = 0.0048, Welch's t test). (ii) Percentage of time spent in the surrounding zone, N = 20 for the WT and N = 22 for the DAT^{PI4KIIIβ−/−} mice (t₍₄₀₎ = 0.9045, p = 0.3711, unpaired t test). I, TST of male mice. Immobility time, N = 20 for the WT mice and N = 22 for the DAT^{PI4KIIIβ−/−} mice (t₍₄₀₎ = 5.681, ****p < 0.0001, unpaired t test). J, Elevated plus mazes of male mice. The time spent in the open arms, N = 20 for the WT mice and N = 22 for the DAT^{PI4KIIIβ−/−} mice (U = 216.5, p = 0.9354, Mann–Whitney U test). K, OFT of female mice. (i) The total distance traveled, N = 8 for the WT mice and N = 8 for the DAT^{PI4KIIIβ−/−} mice (t_(8.86) = 3.83, **p = 0.0041, Welch's t test). (ii) Percentage of time spent in the surrounding zone, N = 8 for the WT and N = 8 for the DAT^{PI4KIIIβ−/−} mice (t₍₁₄₎ = 1.521, p = 0.1505, unpaired t test). L, TST of female mice. Immobility time, N = 8 for the WT mice and N = 8 for the DAT^{PI4KIIIβ−/−} mice (t₍₁₄₎ = 3.522, **p = 0.0034, unpaired t test). M, Elevated plus mazes of female mice. The time spent in the open arms, N = 8 for the WT mice and N = 8 for the DAT^{PI4KIIIβ−/−} mice (t₍₁₄₎ = 2.066, p = 0.0578, unpaired t test).