Figure 2.

Intracellular Ca²⁺ of ALA gradually increases during motion bouts

(A) Schematic of SM and MS transitions. Gray areas represent sleep bouts.

(B) The distribution of the durations of motion bouts (left) and sleep bouts (right) in AML70 lite-1(ce314); wtfIs5[Prab-3::NLS::GcaMP6s; Prab-3::NLS::tagRFP]. The dashed rectangles indicate the bouts that were targeted in the subsequent ALA activity analyses.

(C) Individual (light blue or red) and averaged (dark blue or red) dynamics of ALA intracellular Ca²⁺ during motion bouts (left) or sleep bouts (right) in AML70 lite-1(ce314); wtfIs5[Prab-3::NLS::GcaMP6s; Prab-3::NLS::tagRFP]. The locomotor activity is also shown for motion bouts (left lower panel). Gray areas represent sleep bouts.

(D) Averaged dynamics of intracellular Ca²⁺ and locomotor activity along motion bouts (left) or sleep bouts (right). Here, the duration of each sleep or motion bout was normalized to 1. Shaded areas indicate ±standard error of the mean (SEM). p-values in Wilcoxon signed-rank test are indicated.

(E) Averaged dynamics of Ca²⁺ activity around SM and MS transitions. Shaded areas indicate ±SEM. p-values in paired t test are indicated. Gray areas represent sleep bouts.

(F) Expression pattern of mCherry in SLP984 remEx396[Plet-23::GCaMP6s, Plet-23::mCherry].

(G) The distribution of the durations of motion bouts (left) and sleep bouts (right) in SLP984 remEx396[Plet-23::GCaMP6s, Plet-23::mCherry]. The dashed rectangles indicate the bouts that were targeted in the subsequent ALA activity analyses.

(H) Individual (light blue or red) and averaged (dark blue or red) dynamics of intracellular Ca²⁺ during motion bouts (left) and sleep bouts (right) measured in SLP984 remEx396[Plet-23::GCaMP6s, Plet-23::mCherry]. Gray areas represent sleep bouts.

(I) Averaged dynamics of Ca²⁺ activity measured in SLP984 remEx396[Plet-23::GCaMP6s, Plet-23::mCherry]. Here, the duration of each sleep or motion bout was normalized to 1. Shaded areas indicate ±SEM. p-values in Wilcoxon signed-rank test are indicated.

(J) Average dynamics of ALA Ca²⁺ activity around SM and MS transitions in SLP984 remEx396[Plet-23::GCaMP6s, Plet-23::mCherry]. Shaded areas indicate ±SEM. p-values in Wilcoxon signed-rank test are indicated. Gray areas represent sleep bouts.

(K) Correlation between the latency to peak Ca²⁺ activity and the intensity of peak Ca²⁺ activity during motion bouts. Corresponding p-values are the probabilities of obtaining the observed correlation by chance, when the true correlation is zero. The thick dotted line represents the linear regression line and thin dotted lines show the upper and lower limits of the 95% confidential interval for the regression line. N and n in the figure represent numbers of animals and bouts, respectively.