Dopamine enhances glutamatergic transmission in the dlBNST
A) Diagram of a coronal slice adapted from mouse brain atlas outlining the position of the region of interest, the dlBNST. Immunofluorescent image demonstrates the presence of both TH+ (green) fibers and CRF+ (red) neurons within this region.
B) Representative sEPSC recordings in the dlBNST demonstrating the ability of dopamine to enhance glutamatergic transmission. Scale bars represent 25 pA and 250 ms.
C) A brief (5 minute) application of 1 μM dopamine transiently increases sEPSC frequency in the dlBNST.
D) A brief (5 minute) application of 1 μM dopamine transiently increases sEPSC amplitude in the dlBNST. Inset shows representative normalized sEPSC traces, demonstrating the lack of effect on the kinetics of the response. Scale bar represents 5 ms.
E) Bar graph demonstrating the concentration-dependent effects of dopamine on sEPSC frequency in the dlBNST.
F) Bar Graph demonstrating the effects of multiple concentrations of dopamine on sEPSC amplitude in the dlBNST.

Activation of both D1-like and D2-like receptors is required for dopamine enhancement of spontaneous glutamatergic transmission in the dlBNST
A) A brief application of dopamine (1μM) does not alter sEPSC frequency in the presence of either the D1-like receptor antagonist SCH23390 or the D2-like receptor antagonist sulpiride.
B) Bar graph demonstrating dopamine does not enhance sEPSC amplitude in the presence of the D1-like receptor antagonist SCH23390 and the D2-like receptor antagonist sulpiride or in slices obtained from D1R knockout mice.
C) A brief application of dopamine (1μM) does not alter sEPSC amplitude in the presence of either the D1-like receptor antagonist SCH23390 or the D2-like receptor antagonist sulpiride.
D) Bar graph demonstrating dopamine does not enhance sEPSC amplitude in the presence of the D1-like receptor antagonist SCH23390 and the D2-like receptor antagonist sulpiride or in slices obtained from D1R knockout mice.
E) A brief (5 minute) application of 1 μM dopamine does not alter the frequency of mEPSCs in the BNST.
F) A brief (5 minute) application of 1 μM dopamine does not alter the amplitude of mEPSCs in the BNST.

Dopamine increases excitability in a subpopulation of neurons in the BNST.
A) Representative current-clamp recording from a neuron in the dBNST that was depolarized by dopamine. Each individual trace reflects a current injection ranging from −30 to 40 pA with a 10 pA interval. Scale bars represent 200 ms and 20 mV.
B) Scatter plot demonstrating that dopamine application resulted in a depolarization in a subpopulation of neurons. Neurons that were dramatically depolarized by dopamine are highlighted by a dashed box.
C) Representative recording in a dopamine-responsive neuron demonstrating the depolarizing shift following bath application of 1 μM dopamine. Scale bars represent 1 minute and 5 mV.

CRF signaling is required for dopamine modulation of glutamatergic transmission in the BNST.
A) Close up of a CRF positive neuron (red) within the dlBNST in which TH positive puncta (green) are localized on to the soma. The scale bar is 20 μm. (See supplemental methods for details)
B) Bath application of the CRF-R1 antagonist, NBI 27914 (1 μM), blocks the ability of 1 μM dopamine to enhance sEPSC frequency in the dlBNST.
C) A brief (5 minute) application of 300 nM CRF enhances sEPSC frequency in the dlBNST
D) A brief (5 minute) application of 300 nM CRF does not alter sEPSC amplitude in the dlBNST.
E) Bar graph demonstrating the concentration-dependent effects of CRF on sEPSC frequency in the dlBNST
F) Bar graph demonstrating the effect of a range of concentrations of CRF on sEPSC amplitude
G) A 10 minute application of 300 nM Urocortin 1 enhances sEPSC frequency in the dlBNST
H) A 10 minute application of 300 nM Urocortin 1 does not alter sEPSC amplitude in the dlBNST.

CRF acts through CRF-R1 receptors to enhance glutamate release in the dlBNST
A) CRF induced increase in sEPSC frequency is blocked by pre-application of the CRF-R1 selective antagonist, NBI 27914, but persists in the presence of the CRF-R2 selective antagonist, Astressin-2B
B) Bar graph demonstrating the effects of CRF-R1 and CRF-R2 antagonism on the ability of 300 nM CRF to alter sEPSC frequency.
C) CRF does not alter sEPSC amplitude in the presence of either NBI27914 or Astressin-2B.
D) Bar graph demonstrating the effects of CRF-R1 and CRF-R2 antagonism on the ability of 300 nM CRF to alter sEPSC amplitude.
E) Bath application of 300 nM CRF results in a significant increase in mEPSC frequency as denoted by the shift in the cumulative probability distribution of the interevent interval. Inset: bar graph demonstrating the average normalized increase in frequency (* p < 0.05)
F) Bath application of 300 nM CRF does not result in a significant increase in mEPSC amplitude as denoted by the lack of a shift in the cumulative probability distribution of mEPSC amplitude.

Cocaine produces an enhancement of NMDAR-dependent plasticity following tetanization in the dlBNST.
A) Synaptic potentiation following tetanization (two 100Hz 1 sec trains with a 20 sec interstimulus interval) in mice receiving an acute injection of cocaine (20 mg/kg) or saline 30 minutes prior to sacrifice. Inset, representative traces 10 minutes after the tetanus depicting the enhancement of the N2 portion.
B) Cocaine (3μm) was bath applied for 30 minutes followed by a 60 minute washout before tetanus. Following tetanization enhanced potentiation was observed. Time-matched controls in which no drug was applied to the slice (n=4) are included in the time-course from minutes 0 to 100.
C) Bath application of 100μm APV for 30 minutes before the tetanus attenuates LTP both when cocaine was pre-applied to slices and in slices naïve to cocaine application.
D) Quantification of effects in panels C and D (0–20 minutes post tetanus, ** p<0.01).

Cocaine-induced enhancement of plasticity is dependent on dopamine and CRFR1 signaling
A) Diagrammatic representation of the time-course of experiments shown in this figure. The arrow indicates tetanization. i 10 nM GBR12909 was bath applied for 30 minutes followed by a 60 minute wash-out prior to tetanization. ii 10 μM 10μm flupenthixol was applied 30 minutes prior to cocaine followed by co-application with cocaine. Antagonist was removed 10 minutes following the removal of cocaine and tetanization was performed 50 minutes following removal of antagonist. iii 1μm NBI27914 was applied 30 minutes prior to cocaine followed by co-application with cocaine. Antagonist was removed 10 minutes following the removal of cocaine and tetanization was performed 50 minutes following removal of antagonist.
B) Following tetanization, enhanced short-term potentiation was observed following exposure to GBR12909.
C) Following application of cocaine in the presence of the pan-dopamine receptor antagonist, flupenthixol, there was no alteration in plasticity following tetanus.
D) Following application of cocaine in the presence of the CRFR1 antagonist, NBI27914, there was no alteration in plasticity following tetanus.
E) Quantification of effects of flupenthixol and NBI-27914 on alterations in tetanus evoked plasticity following cocaine exposure (0–20 minutes post tetanus, *p<0.05).
F) Synaptic potentiation following tetanization (two 100Hz 1 sec trains with a 20 sec interstimulus interval) in D1R KO mice receiving an acute injection of cocaine (20 mg/kg) or saline 30 minutes prior to sacrifice.