Input resistances and time constants increase, and excitability also rises. The loss of functional Cv-c from dorsal FB neurons locks the cells in a high-conductance state that likely corresponds to one extreme of the normal operating range of the sleep homeostat (Figure 7). The inability of mutants to exit this high-conductance state despite intense buy Forskolin sleep pressure (Figures 2 and 7) suggests that an essential role of Cv-c is to tune the channel repertoire of sleep-control neurons. Some of the putative substrates of Cv-c, small GTPases of the Rho family (Denholm et al., 2005), have indeed been implicated in various forms of ion channel regulation. RhoA in

its active, GTP-bound, membrane-associated state modulates the conductances of delayed rectifier potassium currents (Cachero et al., 1998). Rac1 in its active state promotes the fusion of vesicles containing transient receptor potential channels and thereby increases channel densities in the plasma membrane (Bezzerides et al., 2004). These precedents illustrate the wide range of potential small GTPase substrates, cellular processes, and ion channel targets that future work will have to sift through in order to arrive at a complete molecular description

of the sleep homeostat. That said, there still remains a formal possibility that the function of Cv-c in sleep control might be divorced altogether from its catalytic Metalloexopeptidase role in the guanine nucleotide cycle of Rho family proteins. Intriguingly, independent evidence already points to the importance of ion channels in sleep control. Candidate see more genes identified in mutagenesis or small-molecule screens encode the fast delayed rectifier potassium channel Shaker ( Cirelli et al., 2005) as well as its cytoplasmic beta

subunit hyperkinetic ( Bushey et al., 2007) and its extracellular regulator sleepless (or quiver) ( Koh et al., 2008), the slow delayed rectifier potassium channel ether-à-go-go ( Rihel et al., 2010), and the voltage-gated sodium channel narrow abdomen ( Lear et al., 2005). Our discovery that ion channel modulation in sleep-control neurons lies at the core of sleep homeostasis offers a physiological context for the pursuit of these leads. Fly stocks were grown on standard media of sucrose, yeast, molasses, and agar and maintained on a 12 hr light/12 hr dark schedule. The following strains were used: cv-cMB03717, cv-cMB01956, cv-cDG20401 ( Bellen et al., 2011 and Venken et al., 2011); cv-cC524, UAS–cv-c ( Denholm et al., 2005); UAS–cv-cRNAi ( Billuart et al., 2001); UAS–CD8-GFP ( Lee and Luo, 1999); C5–GAL4 ( Yang et al., 1995); 104y–GAL4 ( Rodan et al., 2002 and Sakai and Kitamoto, 2006); C205-GAL4 ( Martin et al., 1999); 23E10–GAL4 ( Jenett et al., 2012); tubP–GAL80ts ( McGuire et al., 2003). Baseline sleep was measured as previously described (Shaw et al., 2002).