Figure 6

(A) Schematic showing how A??Prp interactions signal through mGluR5 to activate Fyn kinase, leading to synaptic changes (Nygaard et al., 2014). Small molecules that block each step in the pathway are indicated. (B) Representative traces of sleep behavior after A?long versus A?rev injections in the absence (left) or presence (right) of the A?-Prion binding disruptor, Chicago Sky Blue 6B (3 nM). Ribbons represent ± SEM. (C) The effect of A?long relative to A?rev on normalized sleep during the first day in the in the absence or presence of 3 nM Chicago Sky Blue 6B. The data is pooled from n = 2 independent experiments **p?0.01, one-way ANOVA. (D) Representative traces of sleep behavior after A?long versus A?rev injections in the presence of mGluR5 inhibitor MPEP (5 uM, left) and Fyn Kinase inhibitor saracatinib (300 nM, right). Ribbons represent ± SEM. (E) The effect of A?long relative to A?rev on normalized sleep during the first day in the absence or presence of 5 uM MPEP (left) and 300 nM saracatinib (right). Each dot represents a single larva normalized to the mean A?rev. Data is pooled from two independent experiments. **p?0.01, ****p?10?5 one-way ANOVA. (F) The effect of a 1:1 mixture of A?long to A?short relative to single injections of A?rev, A?short, and A?long on normalized sleep during the first day. The data is pooled from n = 4 independent experiments. (G) A bi-directional model for sleep/wake regulation by A?. In wild-type animals (centre), injection of A?short species signal through Adrb2a/Pgrmc1 to drive wakefulness while A?long oligomers signal via Prp to induce sleep. In mutants that lack Prp (left), only A?short species (as shown by the overlapping distributions) remain to inhibit sleep with no residual A?long oligomers to stimulate the sleep-inducing pathway to counteract wake-inducing signals. Thus prp1-/-; prp2-/- mutants have enhanced wakefulness in response to A?. Conversely, mutants that lack Adrb2a/Pgrmc1 (right), retain only the sleep-promoting A? pathway and fail to increase wakefulness in response to A?short. See also Figure 6?figure supplement 1.