Fig. 3 Imaging DA release in the presence of DA receptor agonists and antagonists. Imaging DA release in the presence of DA receptor agonists and antagonists.

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Fig. 3 Imaging DA release in the presence of DA receptor agonists and antagonists. Imaging DA release in the presence of DA receptor agonists and antagonists. (A) In vitro solution phase maximal ∆F/F (amplitude change at ~1128 nm) of nIRCat in the presence of 100 µM DA; the D2R antagonists sulpiride and haloperidol; D2R agonist quinpirole, and D1R antagonist SCH 23390; and drugs and DA. The addition of 1 μM drug quantities did not induce nIRCat fluorescence modulation in the absence of DA (****P < 0.0001 compared to DA ∆F/F). Subsequent addition of DA to drug-incubated nIRCat solutions produced ∆F/F responses indistinguishable from DA-only responses. Error bars represent SDs from n = 3 measurements. n.s., not significant. (B) Top: A schematic of the effect of D2R agonist and antagonist drugs on DA release. Bottom: Quinpirole suppressed nIRCat fluorescence modulation (****P < 0.0001), whereas sulpiride facilitated nIRCat fluorescence (***P = 0.001) in n = 3 biological replicates. Individual data point represents (∆F/F)max ratio of the average trace collected in same field of view (post-/pre-drug application). (C and D) In brain slice, quinpirole (1 μM) suppressed nIRCat fluorescence modulation in response to a single-pulse electrical stimulation (0.5 mA, 3 ms; red trace) compared to pre-drug ACSF (black trace) but recovered following drug washout (purple and orange traces). (E and F) Sulpiride (1 μM) enhanced nIRCat fluorescence modulation in response to single-pulse electrical stimulation, yielding brighter nIRCat ∆F/F hotspots compared to drug-free ACSF. Scale bars, 10 μm. All error bands in (C) and (E) represent SD from the mean trace. Abraham G. Beyene et al. Sci Adv 2019;5:eaaw3108 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).