Extended Data Fig. 8: Ratiometric calcium imaging of ellipsoid-body-projecting dopaminergic neurons during delay conditioning.
From: Differential mechanisms underlie trace and delay conditioning in Drosophila
Confocal fluorescence images of a, TH-Gal4>>UAS-myr-EGFP, b, MB504b-Gal4>>UAS-myr-EGFP, c, c346-Gal4>>UAS-myr-EGFP, and d, c346-Gal4, TH-FLP>>UAS-FRT-stop-FRT-mCD8-GFP expression in female Drosophila brains staining the pattern of dopaminergic neurons in most of the brain (does not include the PAM cluster), PPL1 dopaminergic cluster projecting to the mushroom bodies, and PPM3 dopaminergic cluster projecting to the ellipsoid body, respectively. In a, left and right images correspond to different z-planes across the brain highlighting dopaminergic neuron subsets targeting the central complex and mushroom body structures. Scale bar is 50 μm. At least three brains were immunostained per genotype presented in a–d. e, Ratiometric imaging of a c346-Gal4, UAS-GCaMP6m.myr-tdTomato female during a single delay conditioning experiment without (air puff) distractors (left), and with distractors (right). Shown, raw dFratio/Fratio activity traces (where Fratio = FGCaMP6m/FtdTomato) for each training trial. f, Peak dFratio/Fratio activity of ellipsoid body DANs during CS (left), and post-US (right), across trials, without distractors (blue, n = 5 flies), and with distractors (red, n = 5 flies). Box plot centre (median), edges (IQR), whiskers (1.5x IQR). Scatters represent individual fly activity scores. Group comparisons were performed using a two-factor ART-ANOVA test. No significant difference was observed between groups.
