Figure 6. Onset dynamics of activity in multiple functional components across the brain at start of walk.

Walk-onset triggered average activity for pan-neuronal data in individual active components at 30 Hz or higher temporal resolution. Trials were normalized individually before averaging (see Figure 6—figure supplement 1 for non-normalized traces and all recording frame rates). Red band indicates walk onset. Note that while most components are activated after walk onset, several components show activity before start of walk. Stars correspond to an above zero significance for integrated traces from −0.5 s to start of walk (Wilcoxon one-sided test). Multiple comparison adjusted p-values (Benjamini–Hochberg): *p < 0.05, **p < 0.01, ***p < 0.001. See Table 2 for definition of acronyms. N = 30 flies of different genotypes, see table in methods for details.

Figure 6—figure supplement 1. Graphs show onset of walk trials for all flies and all transgenic lines at all recording speeds.

The gray band represents the uncertainty of walk-onset time due to lower temporal resolution in some recordings. A corrective factor based on regression coefficient with walk (fluorescence response to walk = coeff × UAS-GCaMP version: GCaMP7f: 0.6, GCaMP6f: 1, GCaMP6M: 1.1, GCaMP7s: 1.2, GCaMP6s: 1.4) was applied for direct comparison between experiments with different reporter versions. The average value for the first 50 ms was taken as baseline and subtracted. Gray shaded area represents walk onset.

Figure 6—figure supplement 2. Normalized trials ∆F/F GCaMP fluorescence in brain regions 1 s before to 1 s after walk onset for different anatomically defined brain regions: (A) dopaminergic neurons (TH/DDC-Gal4;UAS-GCaMP), (B) octopaminergic neurons (Tdc2-Gal4;UAS-GCaMP), and (C) serotonergic neurons (Trh-Gal4;UAS-GCaMP).

Gray shading corresponds to the standard deviation. Red band is the onset of walk.