Extended Data Fig. 6: Monitoring in vivo ACh release induced by electrical stimulation in Drosophila.
From: An optimized acetylcholine sensor for monitoring in vivo cholinergic activity
a, Schematic illustration depicting the experiment in which a transgenic fly expressing ACh3.0 in the KC cells in the mushroom body is placed under a two-photon microscope, and a glass electrode is placed near the mushroom body and used to deliver electrical stimuli. The fly brain is bathed in AHLS containing 100 μM nicotinic acetylcholine receptor blocker mecamylamine (Meca). b, Pseudocolor images (top) and representative traces (bottom) of the fluorescence change in ACh3.0 in response to 2 s of electrical stimulation at the indicated frequencies. Where indicated, the M3R antagonist tiotropium (Tio, 10 μM) is applied to the bath solution. Similar results as the representative images were observed for 8 flies. c, Group summary of the data shown in panel (b); n = 8 flies, p=0.0004. d, ACh3.0 fluorescence is measured before and after a 200-ms electrical stimulation, and the rise and decay phases are fitted with a single-exponential function; the time constants are indicated and summarized on the right; n = 3 flies. All data are shown as mean value ± SEM, with the error bars or shaded regions indicating SEM. Two-sides Student’s t test performed in (c); ***p<0.001.
