Figure 3. GABA and NPY decrease GABAergic tone onto PVH^Crh neurons.

a, Schematic of evoked-IPSC (eIPSCs) recordings from PVH^Crh neurons. b, Example trace of evoked current amplitude from PVH^Crh neurons after Vehicle or NPY treatment. c, NPY reduced eIPSC amplitude in PVH^Crh neurons (Vehicle: 3 neurons/2 mice; NPY: 6 neurons/3 mice). d, Mean eIPSC amplitude in PVH^Crh neurons upon drug application (Two-tailed paired t-test- **P = 0.0017). e, NPY2R blocker prevents NPY-mediated reduction in eIPSC amplitude in PVH^Crh neurons (NPY1/2/5R blocker: 6 neurons/4 mice; NPY1/5R blocker: 9 neurons/4 mice; NPY2R blocker: 7 neurons/4 mice). f, Mean eIPSC amplitude in PVH^Crh neurons after NPY + Blockers (NPY1/2/5R Two-tailed unpaired t-test- P = 0.6353; NPY1/5R Two-tailed unpaired t-test- *P = 0.0216; NPY2R Two-tailed unpaired t-test- P = 0.6353). g, Baclofen reduced eIPSC amplitude in PVH^Crh neurons (7 neurons/6 mice). h, Mean eIPSC amplitude in PVH^Crh neurons after Baclofen (Two-tailed paired t-test- ***P = 0.0004). i, GABA-B-R antagonist prevents baclofen-mediated reduction in eIPSC amplitude in PVH^Crh neurons (4 neurons/2 mice). j, k, Paired-pulse ratio increased in PVHCrh neurons after NPY (j) (6 neurons/3 mice. Two-tailed paired t-test- **P = 0.0015) Baclofen (7 neurons/6 mice. Two-tailed paired t-test- ***P = <0.0001) (k). l, m, Input resistance was unchanged after NPY (l) (6 neurons/3 mice. Two-tailed paired t-test- P = 0.6662) or Baclofen (m) (7 neurons/3 mice. Two-tailed paired t-test- P = 0.7046). n, Time-course of eIPSC amplitude in PVHCrh neurons upon application of Baclofen then NPY (7 neurons/6 mice). o, Time-course of eIPSC amplitude in PVHCrh neurons upon application of NPY then Baclofen (5 cells/3 mice). p, Mean eIPSC amplitude in PVHCrh neurons after application of NPY, then Baclofen. (One-way ANOVA; Tukey’s multiple comparisons. Baseline vs. NPY, *P = 0.0315; Baseline vs.+NPY+Baclofen- ***P = 0.0002; +NPY vs. +NPY+Baclofen- *P = 0.0256).