Figure Legends
Fig. 1. Experimental design. Rats received surgery of implantation of cannula into PVN. The rats were exposed to the vehicle or CORT via drinking water for 21 days from day 1 to day 22. The rats received intra-PVN injections of RU486 and spironolactone, alone or combined from day 16 to day 22. The rats underwent behavioral experiments including novelty-suppressed feeding test, social interaction test, forced swim test and sucrose preference test on day 22 and day 23. The rats were decapitated after the behavioral tests on day 23.
Fig. 2. Chronic CORT treatment for 21 days induced depressive- and anxiety-like behaviors, decreased GR and MR protein levels, and increased CRF protein levels in the PVN. Immobility time in the FST (A) and sucrose preference in the SPT (B) were detected to evaluate depressive-like behaviors. Social interaction in the SIT (C) and latency to feed in the NSFT (D) were detected to evaluate anxiety-like behaviors (n1 , n2 = 9). (E-I) Quantification of MRs (E), GRs (F), the MR/GR ratio (G), and CRF (H) in the PVN and representative Western blots (I) after 21 days of vehicle or CORT administration. β-actin is shown as a quantitative loading control (n1 , n2 = 5). The data are expressed as mean ± SEM. *p < 0.05, **p< 0.01, vs . vehicle group (unpaired Student’st -test).
Fig. 3. Intra-PVN administration of RU486 for 7 days before the test had no effect on depressive- or anxiety-like behaviors in rats that were chronically exposed to CORT via drinking water for 21 days. Immobility time in the FST (A) and sucrose preference in the SPT (B) were detected to evaluate depressive-like behaviors. Social interaction in the SIT (C) and latency to feed in the NSFT (D) were detected to evaluate anxiety-like behaviors (n1 ,n2 , n3 = 6,n4 = 9). (E-I) Quantification of MRs (E), GRs (F), the MR/GR ratio (G), and CRF (H) in the PVN and representative Western blots (I). β-actin is shown as a quantitative loading control (n1 , n2 ,n3 , n4 = 5). The data are expressed as mean ± SEM. *p < 0.05, **p< 0.01, vs . vehicle group; #p< 0.05, ##p < 0.01,vs . CORT alone group (one-way ANOVA followed by the Tukey’s post hoc test).
Fig. 4. Intra-PVN infusion of spironolactone (SPL) for 7 days before the test ameliorated chronic CORT-induced anxiety-like behaviors. Immobility time in the FST (A) and sucrose preference in the SPT (B) were detected to evaluate depressive-like behaviors. Social interaction in the SIT (C) and latency to feed in the NSFT (D) were detected to evaluate anxiety-like behaviors (n1 ,n2 , n3 = 6,n4 = 8). (E-I) Quantification of GRs (E), MRs (F), the MR/GR ratio (G), and CRF (H) in the PVN and representative Western blots (I). β-actin is shown as a quantitative loading control (n1 , n2 ,n3 , n4 = 5). The data are expressed as mean ± SEM. *p < 0.05, **p< 0.01, vs . vehicle group; #p< 0.05, ##p < 0.01,vs . CORT alone group (one-way ANOVA followed by the Tukey’s post hoc test).
Fig. 5.Co-administration of RU486 and spironolactone in the PVN produced anxiolytic- and antidepressant-like effects. Immobility time in the FST (A) and sucrose preference in the SPT (B) were detected to evaluate depressive-like behaviors. Social interaction in the SIT (C) and latency to feed in the NSFT (D) were detected to evaluate anxiety-like behaviors (n1 , n2 ,n3 = 6, n4 = 8). (E-I) Quantification of MRs (E), GRs (F), the MR/GR ratio (G), and CRF (H) in the PVN and representative Western blots (I). β-actin is shown as a quantitative loading control (n1 ,n2 , n3 ,n4 = 5). The data are expressed as mean ± SEM. *p < 0.05, **p < 0.01, vs . vehicle group; #p < 0.05,##p < 0.01, vs . CORT alone group (one-way ANOVA followed by the Tukey’s post hoc test).