Figure 7. Silencing PER_in neurons reduces locomotion.

A. Sample walking traces of control, heterozygous E564-Gal4; UAS-Kir2.1, tub-Gal80^ts and homozygous E564-Gal4; UAS-Kir2.1, tub-Gal80^ts flies (3 flies/genotype) without (22 °C) or with Kir2.1 (30°C) induction (top 6 traces). The bottom two traces show mosaic E564 flies selected for either a retracted or extended proboscis. As with the full E564-Gal4 line, flies with extended proboscises moved significantly less. Different colors mark movement paths of different flies for one minute. Scale bar is 10mm.

B. E564-Gal4; UAS-Kir2.1, tub-Gal80^ts flies showed severely reduced movement when compared to controls. This phenotype was also observed in mosaic flies exhibiting constitutive PER, but not in mosaic flies that did not exhibit PER. n=12–15 flies/condition for non-mosaic animals, n=25–30 for mosaics, mean ± SEM, two-way ANOVA for E564-Gal4; UAS-Kir2.1, tub-Gal80^ts heterozygotes versus controls; students t-test for E564-Gal4; UAS-Kir2.1, tub-Gal80^ts homozygotes and mosaics, ***P<0.001.

C. Analysis of mosaic animals selected for high movement (<250mm/minute) or low movement (>200mm/minute). Only cell-type 1 (PER_in) neurons were enriched in flies with low locomotion (cell-types numbered in Figure 2 and S2). n=25–30 flies/condition, mean±SEM, Fisher’s exacttest, ***P<0.001. See Figure S4 for behavioral analyses of a second Gal4 line that contains PER_in.

D. Waxing the proboscis in an extended state, but not in a retracted state, impaired locomotion in wild-type flies. Waxing the proboscis in a retracted state partially rescued the locomotion defects seen upon neural inactivation of E564-Gal4; UAS-Kir2.1, tub-Gal80^ts heterozygous flies. n=12–22 flies/condition for non-mosaic animals, n=23–26 for mosaics, mean ± SEM, two-way ANOVA for E564-Gal4; UAS-Kir2.1, tub-Gal80^ts heterozygous flies versus controls, students t-test for wild-type waxed proboscis,***P<0.001, *P<0.05. See Figure S4, showing that a second Gal4 line, E605-Gal4, containing PER_in shows similar activation and inactivation behavioral phenotypes.