Figure 5
Rogdi-dependent sleep involves a sleep-promoting pathway of GABAergic transmission via metabotropic GABA receptors. (a) A schematic of GABAergic synaptic transmission. DABA, DL-2,4-diaminobutyric acid (GAT inhibitor); EOS, ethanolamine O-sulfate (GABA-T inhibitor); GABAA-R, ionotropic GABA receptor; GABAB-R, metabotropic GABA receptor; GABA-T, GABA transaminase; GAD1, glutamate decarboxylase 1; GAT, GABA transporter; SKF-97541, 3-Aminopropyl(methyl)phosphinic acid (GABAB-R agonist); THIP, 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (GABAA-R agonist); VGAT, vesicular GABA transporter. (b,c) Oral administration of EOS (b) or DABA (c) rescues sleep quantity and sleep latency in Rogdi mutants. Sleep behaviors in individual flies were analyzed similarly to the data presented in Fig. 1. Gray and orange bars indicate wild-type (w 1118) and Rogdi del mutant backgrounds, respectively. Data represent average ± SEM (n = 108–120 for EOS; n = 36–78 for DABA). Two-way ANOVA detected significant interaction of Rogdi mutation with EOS and DABA effects on sleep amount (F[1,449] = 21.43, P < 0.0001 for EOS; F[1,209] = 55.2, P < 0.0001 for DABA), average sleep bout length (ABL) (F[1,449] = 4.284, P = 0.0390 for EOS only) and sleep latency (F[1,449] = 18.61, P < 0.0001 for EOS; F[1,209] = 16.61, P < 0.0001 for DABA). (d) An agonist of GABAB-R (SKF-97541) but not GABAA-R (THIP) fully rescues the short sleep phenotypes in Rogdi mutants (n = 29–92). Two-way ANOVA detected significant interaction of Rogdi mutation with SKF-97541 effects on sleep amount (F[1,228] = 56.39, P < 0.0001), ABL (F[1,228] = 33.81, P < 0.0001) and sleep latency (F[1,228] = 14.38, P = 0.0002) but not with THIP. n.s., not significant, *P < 0.05, **P < 0.01, ***P < 0.001 to no-drug controls in the same genetic backgrounds as determined by Tukey post hoc test.
