Fig. 5 | Molecular Psychiatry

Fig. 5

From: SSRIs target prefrontal to raphe circuits during development modulating synaptic connectivity and emotional behavior

Fig. 5

Bidirectional modulation of developmental SSRI-induced emotional alterations by PFC-SERT+ neurons. a For chemogenetic manipulation of PFC glutamate projection-neurons, AAV5-CaMKIIa-hM4D(Gi)-mCherry (c-e) or AAV8-CaMKIIa-hM3D(Gq)-mCherry (i-k) was bilaterally injected into the PFC of P5 wild-type C57BL/6 mice. For conditional expression in PFC-SERT neurons the AAV5-hSYN-DIO-hM4D(Gi)-mCherry (f-h) and AAV8-hSYN-DIO-hM3D(Gq)-mCherry (l-n) were used in SERTCre/+ mice. Mice were treated with fluoxetine (FLX) (10 mg/kg/day in 5% sucrose) or with 5% sucrose (SUC), from P2 to P14. Behavioral analyses were started at P80, starting with the novelty-suppressed feeding test (NSF), the forced swim test (FST), and locomotor activity spaced by 7 days intervals. b Immobility time in the FST during the first day “drug-free” session. Control group: 25 mice (15 males, 10 females); FLX group: 23 mice (10 males, 13 females). Two-ways ANOVA: gender x treatment interaction (F1,44 = 1.855, p = 0.18); gender main effect (F1,44 = 6.407, p < 0.02); treatment main effect (F1,44 = 5.228, *p < 0.03). c: Immobility time in the FST (day 2); d: latency to feed in the NSF; e: locomotor activity, after a single i.p. injection of saline (NaCl 0.9%) or CNO (1 mg/kg) 30 min. before the testing. Two-ways ANOVA of the FST (c): treatment combination x gender interaction (F3,40 = 1.345, p = 0.27); treatment combination main effect (F3,40 = 9.510, p < 0.0001); gender main effect (F1,40 = 3.550, p = 0.07). FLX-CNO vs. Control-Saline (*p < 0.0001), vs. Control-CNO (*p < 0.001) and vs. FLX-Saline (*p < 0.02) by Tukey’s test. Two-ways ANOVA of NSF (d): treatment combination x gender interaction (F3,40 = 0.657, p = 0.58); treatment combination main effect (F3,40 = 6.994, p < 0.001); gender main effect (F1,40 = 0.011, p = 0.916). FLX-CNO vs. Control-Saline (*p < 0.001), vs. Control-CNO (*p < 0.004) and vs. FLX-Saline (*p < 0.04) by Tukey’s test. Two-ways ANOVA of locomotor (e): treatment combination x gender interaction (F3,40 = 0.245, p = 0.865); treatment combination main effect (F3,40 = 0.394, p = 0.758); gender main effect (F1,40 = 3.318, p = 0.08). In (c-e): Control/Saline group: 13 mice (8 males, 5 females); Control/CNO group: 12 mice (7 males, 5 females); FLX/Saline group: 12 mice (6 males, 6 females); FLX/CNO group: 11 mice (4 males, 7 females). f-h Conditional inhibition of PFC-SERT neurons in SERTCre/+ mice treated or not with fluoxetine (FLX) from P2 to 14) (a). Immobility in the FST (day 2) (f), latency to feed in the NSF (g) and locomotor activity (h) after an i.p. injection of either CNO (1 mg/kg) or NaCl 0.9% saline (SAL), 30 min. before testing. In (f): ANOVA: treatment main effect (F1,9 = 5.982, p < 0.04), in (g): ANOVA: treatment main effect (F1,9 = 6.287, p < 0.04), in (h): ANOVA: treatment main effect (F1,9 = 2.712, p = 0.134). In (f-h): FLX/Saline group: 5 males; FLX/CNO group: 6 males. Immobility time in the FST (day 2) (i), latency to feed in the NSF (j) and locomotor activity (k), after a single i.p. injection of either CNO (1 mg/kg) or NaCl 0.9% saline, 30 min. before testing. Two-ways ANOVA in (i): treatment combination x gender interaction (F3,43 = 0.612, p = 0.61); treatment combination main effect (F3,43 = 4.177, p < 0.02); gender main effect (F1,43 = 0.315, p = 0.58). FLX-Saline vs. control-saline (*p < 0.05), vs. FLX-CNO (*p < 0.01) by Tukey’s test. Two-ways ANOVA in (j): treatment combination x gender interaction (F3,43 = 0.490, p = 0.69); treatment combination main effect (F3,43 = 9.532, p < 0.0001); gender main effect (F1,43 = 3.218, p = 0.08). FLX-Saline vs. Control-Saline (*p < 0.0002), vs. Control-CNO (*p < 0.0001) and vs. FLX-CNO (*p < 0.01) by Tukey’s test. Two-ways ANOVA in (k): treatment combination x gender interaction (F3,43 = 0.815, p = 0.492); treatment combination main effect (F3,43 = 0.521, p = 0.670); gender main effect (F1,43 = 0.137, p = 0.713). In (i-k): Control/Saline group: 16 mice (8 males, 8 females); Control/CNO group: 12 mice (8 males, 4 females); FLX/Saline group: 12 mice (6 males, 6 females); FLX/CNO group: 11 mice (4 males, 7 females). l-n Conditional activation of PFC-SERT neurons in SERTCre/+ mice treated or not with fluoxetine (FLX) from P2 to P14). a Immobility in the FST (day 2) (l), latency to feed in the NSF (m) and locomotor activity (n) after an i.p. injection of either CNO (1 mg/kg) or NaCl 0.9% saline (SAL), 30 min. before the testing. In (l): ANOVA: treatment main effect (F1,13 = 5.100, p < 0.05), in (m): ANOVA: treatment main effect (F1,13 = 13.792, p < 0.01), in (n): ANOVA: treatment main effect (F1,13 = 0.0006, p = 0.981). In (l-n): FLX/Saline group: 8 males; FLX/CNO group: 7 males

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