Fig. 5: Recording from Mooncrawler Descending Neuron during forward and backward crawling.
From: Closed-loop two-photon functional imaging in a freely moving animal
One MDN cell body was tracked to stabilize imaging of a larger (150 × 150 × 50 μm) volume. The tracked cell body location was extracted from this larger volume and used to quantify fluorescence ratios. a Left: Ratiometric activity measure (green fluorscence/red fluorescence) vs. time for eight different larvae expressing mCherry and GCaMP6f in MDN. Behavioral state (green = forward, red = backward, and black = other) is indicated by color. Each trace is normalized so that the ratiometric measure averaged over all times the larva was crawling forward is 1. Right: Box and whisker plot for each experiment showing the activity measure for each behavioral bout (continuous period of forward or backward crawling) separated by forward and backward crawling. Each data point represents the average ratiometric measure over a single behavioral bout. Central line is the median, box spans 25th–75th percentiles. */** rejects the null-hypothesis that the median of all forward crawling bouts is equal to or greater than the median of all backward crawling bouts (one-sided Wilcoxon rank-sum test) at p < 0.05/0.01, respectively (p-values: i:7.9e-06, ii:0.84, iii:0.25, iv:0.0047, v:0.022, vi:0.047, vii:0.01, viii:0.0023). c Same as b, except MDN was labeled with mCherry and Calcium insensitive GFP. (p-values ix:0.33, x:0.04, xi:0.94, xii:0.67, xiii:0.18, xiv:0.94, xv:0.47). c Box-and-whisker plot of the activity measure of each bout shown in (a, b) separated by behavioral state and indicator. Each bout is shown as a small dot; bouts from the same experiment are aligned vertically. Central line is the median of all bouts, box spans 25th–75th percentiles, and whiskers span the 5th–95th percentiles. The hypothesis that the GCaMP forward median is greater than or equal to the GCaMP backward median is rejected at p = 3 × 10−9 by the one-sided Wilcoxon rank-sum test. The hypothesis that the GCaMP backward median is the same as the GFP backward median is rejected at p = 8 × 10–7 by the two-sided Wilcoxon rank-sum test.
