Mechanisms of Circadian Clock Output

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DESCRIPTION (provided by applicant): We are interested in signaling mechanisms used by circadian pacemaker neurons to organize daily locomotor behavior. There has been tremendous progress in recent years in defining the molecular basis of the cell autonomous clockwork mechanism that underlies circadian timekeeping. That definition has permitted the identification of the primary circadian clock neurons within the brain. In turn, that has presented the opportunity to re-examine fundamental issues concerning the neural basis of behavior. This research program features cellular, molecular and genetic techniques to understand how signals are passed from the primary pacemakers to responsive neurons. Previously we showed that the neuropeptide PDF is the principal circadian transmitter in Drosophila. Genetic evidence indicates it is the most important signal used by the primary clock cells (the LN-v neurons). We will extend that information by performing experiments that are designed to ask where and when PDF signals circadian time within the brain. Further, we will identify and characterize the receptor(s) for PDF. Definition of the PDF receptor will permit a description of which neural elements and which circuits respond to the circadian transmitter. We can also begin to ask what are the long-term consequences (signal transduction pathways) that are modulated as a result of PDF receptor activation. Because circadian signaling occurs on such a long time base (many hours), we hypothesize that PDF signaling may include transcriptional regulation. We have identified several genes whose expression levels are pdf-dependent and which cycle on a daily basis. We propose now to demonstrate their role as PDF targets for the mediation of circadian signaling. Finally, we have performed an extensive genome-wide microarray screen for circadian gene expression in the Drosophila head. We have identified 22 circadian genes, of which 15 are novel in the context of circadian biology. We hypothesize that most of these circadian genes participate in circadian output mechanisms, and propose experiments to address this possibility