Extended Data Fig. 6: Ratiometric calcium imaging of ellipsoid body ring neurons during delay conditioning reveals an increased oscillatory component and steady decline with repeated training that is not susceptible to distractions.
From: Differential mechanisms underlie trace and delay conditioning in Drosophila
a, Ratiometric imaging of an EB1-Gal4, UAS-GCaMP6f.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 = FGCaMP6f/FtdTomato) for trials 1, 4, and 7. Red curve fits are single-term exponential model fits through the raw dFratio/Fratio activity traces starting at the end of CS. b, Scree plots of cumulative variance explained by each of the top 50 principal components for all delay conditioning trials without distractors (blue), and with distractors (red). c, Cumulative variance explained by the top 12 components per trial for delay conditioning without distractors (blue), and with distractors (red). d, Exponential curve fit decay rates of EB1 neural activity post-CS for trials without distractors (blue), and with distractors (red). e, Normalized relative power of EB1 neural activity for trials 1, 4 and 7 (left to right) during CS. Shown are the detected frequencies at maximum relative power for trials without distractors (blue), and with distractors (red), greater than peak mean baseline frequency. Black line represents mean baseline frequency spectrum (from n = 51 flies). f, Frequency (at maximum relative power) of EB1 neural activity across delay conditioning trials during CS for trials without distractors (blue), and with distractors (red) (see Methods, ‘Frequency and decay rate computation’). No distractor trials (n = 9 flies), distractor trials (n = 9 flies). g, Frequency (from f) as a function of learning performance (see Methods, ‘Correlating physiological and behavioural metrics’) for trace conditioning with (red, R2 0.72) and without (blue, R2 0.77) distractors. Curve fits are second degree polynomials through the median activity for each of the no distractor (blue) and distractor (red) trials. 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.
