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Fig. 4 | Molecular Brain

Fig. 4

From: A novel red fluorescence dopamine biosensor selectively detects dopamine in the presence of norepinephrine in vitro

Fig. 4

Characterization of the red-DA biosensor R-GenGAR-DA1.2 in the primary culture of rat hippocampal neurons. a Representative images of a primary hippocampal neuron expressing DA1.2. Top: the fluorescence change (ΔF/F0) before (left) and after the application of 5 μM DA (middle) followed by 5 μM SCH (right) are shown in pseudocolor intensity-modulated display mode. Bottom: magnification of dendrite marked in the top left image (white rectangle). Medium temperature was equilibrated before imaging (Additional file 1: Fig. S5b). b The fluorescence change (ΔF/F0) of DA1.2 in the primary hippocampal neurons in panel (a). DA (5 μM) and SCH (5 μM) were treated at the time points indicated by pink and blue bars, respectively. Mean ΔF/F0 values of 6 neurons from 6 experiments are shown with SD (shaded area). c DA1.2 was pre-treated with SCH before application of DA. Mean ΔF/F0 of 3 neurons from 3 experiments are shown with SD (shaded area). Medium temperature was equilibrated before imaging. d Temperature equilibration dose–response curves of DA (pink) and NE (green) on the primary hippocampal neurons expressing DA1.2 (Additional file 1: Fig. S4d). DA: max ΔF/F0 = − 0.52 ± 0.03 and EC50 = 0.25 ± 0.05 µM; n = 10 neurons from 10 experiments. NE: max ΔF/F0 = − 0.52 ± 0.04 and EC50 = 10 ± 2 µM; n = 7 neurons from 7 experiments. Experimental data (dots) were fitted with the Hill equation (lines). DA1.2 showed 40-fold selectivity for DA over NE in the primary culture of rat hippocampal neurons

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