Fig. 5
The red-DA biosensor R-GenGAR-DA1.2 combined with the green-NE biosensor GRABNE1m enables dual-color fluorescence imaging of DA and NE in a primary culture of rat hippocampal neurons. a Representative images of a primary hippocampal neuron co-expressing DA1.2 and NE1m. b Enlarged time-lapse images of DA1.2 and NE1m treated with agonists or antagonists in the pseudocolor intensity-modulated display mode from the dendritic region in the primary hippocampal neurons marked as the white boxed region in panel (a, left). Concentrations: DA and SCH, 5 µM; NE and YO, 1 µM. Temperature equilibration and pre-illumination were conducted before compound application (Additional file 1: Fig. S5d). c The fluorescence intensity change (ΔF/F0) of DA1.2 (top) and NE1m (bottom) in the primary hippocampal neurons co-expressing DA1.2 and NE1m. Bars show the schedule of agonist/antagonist application to both DA1.2 and NE1m. Gray vertical lines indicate time of application. Vehicle, 10 µM HCl in H2O or 0.1% DMSO; control, cells were only exposed to excitation light. Colored lines indicate mean ΔF/F0 and light-colored shaded area is the SD. Ligands, 6 neurons from 6 experiments; vehicles, 4 neurons from 4 experiments; control, 4 neurons from 1 experiment. Statistical test results are shown in Additional file 1: Fig. S11c, d