Extended Data Fig. 5: Probing endogenous ACh release in mouse brain slices.
From: An optimized acetylcholine sensor for monitoring in vivo cholinergic activity
a–f, Representative fluorescence traces (a, d) and group summary (b, c, e, f) of the fluorescence change (ΔF/F0 and SNR) in neurons expressing either ACh2.0 or ACh3.0 in response to electrical stimulation in MHb-IPN brain slices. The slices are bathed in either ACSF or 2 μM baclofen (Bac). N=5 slices from 3 mice for ACh2.0, and n = 10 slices from 7 mice for ACh3.0. g, h, The representative fluorescence traces (g) and group data (h) of ACh3.0-expressing neurons in response to 100-Hz electrical stimulation with different stimulation times in MHb-IPN brain slices. The response in either ACSF or 100 μM 4-AP is measured and summarized; n = 5 slices from 5 mice. i, The kinetics of fluorescence response of ACh3.0 to a single pulse (2 ms) of electrical stimulation in the presence of 100 μM 4-AP. The response in three independent experiments are normalized and plotted together in the middle. The group data of on and off response time constants are summarized on the right (n = 3 slices from 3 mice). All data are shown as mean value ± SEM, with the error bars or shaded regions indicating SEM.
