Figure 2, supplementary figure 2

Figure 2: BNSTCRF neurons are sensitive to ovarian estrogen status. Spontaneous synaptic transmission in BNSTCRF neurons as measured in slice electrophysiology recordings from intact CRF-CrexAi9 reporter females in low vs. high ovarian E2 states, with schematic in a. b) Representative traces of spontaneous excitatory postsynaptic currents (sEPSCs). c) The frequency of sEPSCs in BNSTCRF neurons from high and low ovarian E2 status females (N’s = 6 low E2, 13 cells; 7 high E2, 13 cells; d and e are the same cells). d) sEPSC amplitude in high and low ovarian E2 status female BNSTCRF neurons. e) Excitatory synaptic drive onto BNSTCRF neurons (frequency x amplitude) high and low ovarian E2 status females. f) Representative traces of spontaneous inhibitory postsynaptic currents (sIPSCs). g-i) sIPSC frequency in BNSTCRF neurons high and low ovarian E2 status females (g), amplitude (h), and synaptic drive (i; N’s = 6 low E2, 12 cells; 7 high E2, 11 cells). *PSupplementary Figure 2: BNSTCRF neurons are necessary for binge alcohol consumption. a-g) Chemogenetic manipulation of BNSTCRF neurons during EtOH Drinking in the Dark (DID) and avoidance behaviors. a-b) Schematic (a) and representative image (b) for viral strategy to bidirectionally manipulate BNSTCRF neurons using a multiplexed Gi+Gq DREADD approach. c-d) Activation of the Gi-coupled KOR DREADD via systemic injection of Salvinorin B (SalB; 10 mg/kg) suppressed binge EtOH consumption compared to vehicle (VEH) injected controls (c; N’s = 7 CON, 5 DREADD) but did not affect the % time in open arms of the elevated plus maze (EPM; left) or distance traveled (right; d; N’s = 6 CON, 5 DREADD). e) DREADD virus hit map in the BNST (for Fig. 2a–f). Each dot is an individual hit and the grey Xs are a miss. f-g) Gq DREADD-mediated BNSTCRF neuron activation via clozapine-N-oxide (CNO; 5 mg/kg i.p.) did not change EtOH consumption (f; N’s = 7 CON, 6 DREADD) or alter avoidance behavior on the open field (OF) measured via % time in center (left) but did reduce distance traveled (right; g; N’s = 14 CON, 9 DREADD).*P