Article Figures & Data
Figures
- Figure 1.
Decreased 5-HT content in the dSTR is correlated with depression-like behaviors. a–c, Results of social interaction test. a, Time in the interaction zone. Naive, n = 9; Resilient, n = 8; Susceptible, n = 8; ***p < 0.005 versus susceptible mice (one-way ANOVA with Bonferroni’s post hoc tests). b, Interaction ratio. Naive, n = 9; Resilient, n = 8; Susceptible, n = 8; ***p < 0.005 versus susceptible mice (one-way ANOVA with Bonferroni’s post hoc tests). c, Time in avoidance zone. Naive, n = 9; Resilient, n = 8; Susceptible, n = 8; ***p < 0.005 versus susceptible mice, **p < 0.01 versus susceptible mice (one-way ANOVA with Bonferroni’s post hoc tests). d, Quantitative of basal 5-HT levels in the dSTR. Naive, n = 7; Resilient, n = 7; Susceptible, n = 6; **p < 0.01 versus susceptible mice, *p < 0.05 versus susceptible mice (one-way ANOVA with Bonferroni’s post hoc tests). e, f, Correlation of basal 5-HT content in the dSTR with the social interaction ability assessed by the time spent in interaction zone (e) and by interaction ratio (f) after RSDS. Resilient, n = 7; Susceptible, n = 6 (Pearson’s correlation test).
- Figure 2.
Pharmacogenetic activation of serotonergic neurons in the dRN. a, Schematic of microinjection of AAV vectors into the dRN and microinfusion of DCZ into the dSTR. b, Representative images of injection sites (dRN). TPH-positive cells: red signal; EYFP-positive cells: green signal. Entire portion (left), Scale bars: 200 μm. Magnified portion (right), Scale bars: 20 μm. c, 5-HT content in dSTR collected every 12 min for 2 h from the time DCZ was microinfused. dRN-Mock with Ringer, n = 4; dRN-Mock with DCZ, n = 5; dRN-hM3Dq with Ringer, n = 5; dRN- hM3Dq with DCZ, n = 5; ***p < 0.005 versus dRN-Mock with Ringer, **p < 0.05 versus dRN-Mock with Ringer (two-way ANOVA with Bonferroni’s post hoc tests).
- Figure 3.
Upregulation of 5-HT content in the dSTR induced resilience to stress in dSTR-Shati OE mice. a, Representative images of injection sites (dSTR). Shati/Nat8l (His-positive cells): green signal, NeuN-positive cells: red signal. Entire portion (right), Scale bars: 500 μm. Magnified portion (left), Scale bars: 50 μm. b, Relative expression level of Shati/Nat8l mRNA in the dSTR. dSTR-Mock, n = 8; dSTR-Shati OE, n = 7; **p < 0.01 versus dSTR-Mock mice (student t test). c, Schematic of microinjection and microinfusion and the timeline of experiments. d–f, Results of social interaction test. d, Time in the interaction zone. dSTR-Mock with ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11; ***p < 0.005 versus dSTR-Mock with Ringer; ###p < 0.005 versus dSTR-Shati OE with DCZ (two-way ANOVA with Bonferroni’s post hoc tests). e, Interaction ratio. dSTR-Mock with Ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11; ***p < 0.005 versus dSTR-Mock with Ringer; ##p < 0.01 versus dSTR-Shati OE with DCZ (two-way ANOVA with Bonferroni’s post hoc tests). f, Time in avoidance zone. dSTR-Mock with Ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11; **p < 0.01 versus dSTR-Mock with Ringer; ###p < 0.005 versus dSTR-Shati OE with DCZ (two-way ANOVA with Bonferroni’s post hoc tests). g, Result of sucrose preference test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11; ***p < 0.005 versus dSTR-Mock with ringer; ##p < 0.01 versus dSTR-Shati OE with DCZ (two-way ANOVA with Bonferroni’s post hoc tests). h, Immobility time in tail suspension test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11; *p < 0.05 versus dSTR-Mock with Ringer (two-way ANOVA with Tukey’s post hoc tests). i, Immobility time in a forced swimming test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11; *p < 0.05 versus dSTR-Mock with Ringer; #p < 0.05 versus dSTR-Shati OE with DCZ (two-way ANOVA with Bonferroni’s post hoc tests). j, Results of locomotor activity test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with DCZ, n = 11; dSTR-Shati OE with Ringer, n = 11; dSTR-Shati OE with DCZ, n = 11 (two-way ANOVA with Bonferroni’s post hoc tests).
- Figure 4.
GABA content in the dRN is correlated with depression-like behaviors and controlled by Shati/Nat8l in the dSTR. a, Quantitative basal GABA content in the dRN. Naive, n = 9; Resilient, n = 8; Susceptible, n = 8; **p < 0.01 versus susceptible mice; *p < 0.05 versus susceptible mice (one-way ANOVA with Bonferroni’s post hoc tests). b, c, Correlation of basal GABA content in the dRN with the social interaction ability assessed by the time spent in interaction zone (b) and by interaction ratio (c) after RSDS. Resilient, n = 8; Susceptible, n = 8 (Pearson’s correlation test). d, Quantitative basal GABA content in the dRN. Mock, n = 8; dSTR-Shati, n = 7; **p < 0.01 versus Mock mice (student t test).
- Figure 5.
Blockade of GABA signaling in the dRN suppressed stress vulnerability in dSTR-Shati OE mice. a, Schematic of microinjection and microinfusion and the timeline of experiments. b–d, Results of social interaction test. b, Time in the interaction zone. dSTR-Mock with ringer, n = 10; dSTR-Mock with CGP36216, n = 11; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9; ***p < 0.005 versus dSTR-Mock with Ringer; ###p < 0.005 versus dSTR-Shati OE with CGP362160 (two-way ANOVA with Bonferroni’s post hoc tests). c, Interaction ratio. dSTR-Mock with Ringer, n = 10; dSTR-Mock with CGP36216, n = 10; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9; ***p < 0.005 versus dSTR-Mock with Ringer; ##p < 0.01 versus dSTR-Shati OE with CGP36216 (two-way ANOVA with Bonferroni’s post hoc tests). d, Time in avoidance zone. dSTR-Mock with Ringer, n = 10; dSTR-Mock with CGP36216, n = 10; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9; **p < 0.01 versus dSTR-Mock with Ringer; ##p < 0.01 versus dSTR-Shati OE with CGP36216 (two-way ANOVA with Bonferroni’s post hoc tests). e, Result of sucrose preference test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with CGP36216, n = 10; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9; ***p < 0.005 versus dSTR-Mock with ringer; ##p < 0.01 versus dSTR-Shati OE with CGP36216 (two-way ANOVA with Bonferroni’s post hoc tests). f, Immobility time in tail suspension test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with CGP36216, n = 10; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9; **p < 0.01 versus dSTR-Mock with Ringer; ###p < 0.005 versus dSTR-Shati OE with CGP36216 (two-way ANOVA with Tukey’s post hoc tests). g, Immobility time in a forced swimming test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with CGP36216, n = 10; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9; **p < 0.01 versus dSTR-Mock with Ringer; ###p < 0.005 versus dSTR-Shati OE with CGP36216 (two-way ANOVA with Tukey’s post hoc tests). h, Results of locomotor activity test. dSTR-Mock with Ringer, n = 10; dSTR-Mock with CGP36216, n = 10; dSTR-Shati OE with Ringer, n = 9; dSTR-Shati OE with CGP36216, n = 9 (two-way ANOVA with Bonferroni’s post hoc tests).
- Figure 6.
Overexpression of Shat/Nat8l enhanced BDNF expression in the dSTR. Relative expression level of Shati/Nat8l mRNA in the dSTR. dSTR-Mock, n = 6; dSTR-Shati OE, n = 5; *p < 0.05 versus dSTR-Mock mice (student t test).
- Figure 7.
Hypothesized mechanism of establishment of stress vulnerability after RSDS. Overview of dSTR-dRN circuitry changes in the mouse brain after RSDS. Shati/Nat8l in the dSTR was increased by RSDS, and BDNF expression in the dSTR and GABA release in the dRN was increased. The serotonergic system in the dRN is deactivated by elevated GABA content, and the 5-HT content in the dSTR is decreased. Finally, stress vulnerability is established, leading to depression-like behaviors in response to stress.
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