Figure 5: GACh sensors reveal dynamics of endogenous ACh release in Drosophila. | Nature Biotechnology

Figure 5: GACh sensors reveal dynamics of endogenous ACh release in Drosophila.

From: A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies

Figure 5

(a) Schematic illustration of the two-photon imaging setup of the Drosophila olfactory system. Odor was delivered near the antenna (left), and GACh signals were measured in the antennal lobe area of GH146-Gal4: UAS-GACh flies (right). (b) Pseudocolor images of GACh-expressing antenna lobes show fluorescence responses to mineral oil and odor isoamyl acetate (IA). (c) Time courses of the IA-dependent responses in DM2 and DA1 glomeruli of GACh1.0- (upper plots) and GACh2.0- (lower plots) expressing antenna lobes. The traces were averaged from three trials in the same fly. (d) Values for the maximal ΔF/F0 in DM2 and DA1 glomeruli of GACh1.0-expressing antenna lobes (upper plot; DM2: 0.29 ± 0.49%, n = 9 flies; DA1: –1.01 ± 0.68%, n = 9 flies, U = 55, P = 0.22 for mineral oil; DM2: 7.02 ± 0.56%, n = 9 flies; DA1: –0.61 ± 0.80%, n = 9 flies, U = 81, P = 4.12 × 10−4 for 10−3 IA; DM2: 12.97 ± 1.28%, n = 7 flies; DA1: 0.17 ± 0.39%, n = 7 flies, U = 49, P = 0.002 for IA 10−2) and GACh2.0-expressing antenna lobes (lower plot; DM2: 2.27 ± 2.34%, n = 8 flies; DA1: –0.73 ± 1.41%, n = 8 flies, U = 35, P = 0.80 for mineral oil, DM2: 18.78 ± 1.36%, n = 10 flies; DA1: –2.37 ± 1.06%, n = 10 flies, U = 100, P = 1.83 × 10−4 for 10−3 IA; DM2: 37.30 ± 4.79%, n = 9 flies; DA1: –0.55 ± 2.68%, n = 9 flies, U = 81, P = 4.12 × 10−4 for 10−2 IA). (e) Upper, IA-evoked responses in DM2 glomerulus of GACh1.0 and GACh2.0 transgenic flies. Lower: values for the maximal ΔF/F0 in DM2 glomerulus of GACh1.0 and GACh2.0 transgenic flies (GACh1.0: 12.97 ± 1.28%, n = 7 flies; GACh2.0: 37.30 ± 4.79% n = 9 flies; U = 63, P = 0.001). Data are shown as mean ± s.e.m.; error bars indicate s.e.m. Experiments in b were repeated independently for more than seven flies with similar results. **P < 0.01 (Mann–Whitney Rank Sum non-parametric tests, two-sided). All scale bars, 10 μm.

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