title: The Drosophila Receptor Protein Tyrosine Phosphatase LAR Is Required for Development of Circadian Pacemaker Neuron Processes That Support Rhythmic Activity in Constant Darkness But Not during Light/Dark Cycles.
abstract: InDrosophila, a transcriptional feedback loop that is activated by CLOCK-CYCLE (CLK-CYC) complexes and repressed by PERIOD-TIMELESS (PER-TIM) complexes keeps circadian time. The timing of CLK-CYC activation and PER-TIM repression is regulated post-translationally, in part through rhythmic phosphorylation of CLK, PER, and TIM. Although kinases that control PER, TIM, and CLK levels, activity, and/or subcellular localization have been identified, less is known about phosphatases that control clock protein dephosphorylation. To identify clock-relevant phosphatases, clock-cell-specific RNAi knockdowns ofDrosophilaphosphatases were screened for altered activity rhythms. One phosphatase that was identified, the receptor protein tyrosine phosphatase leukocyte-antigen-related (LAR), abolished activity rhythms in constant darkness (DD) without disrupting the timekeeping mechanism in brain pacemaker neurons. However, expression of the neuropeptide pigment-dispersing factor (PDF), which mediates pacemaker neuron synchrony and output, is eliminated in the dorsal projections from small ventral lateral (sLNv) pacemaker neurons whenLarexpression is knocked down during development, but not in adults. Loss ofLarfunction eliminates sLNvdorsal projections, but PDF expression persists in sLNvand large ventral lateral neuron cell bodies and their remaining projections. In contrast to the defects in lights-on and lights-off anticipatory activity seen in flies that lack PDF,LarRNAi knockdown flies anticipate the lights-on and lights-off transition normally. Our results demonstrate thatLaris required for sLNvdorsal projection development and suggest that PDF expression in LNvcell bodies and their remaining projections mediate anticipation of the lights-on and lights-off transitions during a light/dark cycle.
  
  
  CONCLUSIONS
  In animals, circadian clocks drive daily rhythms in physiology, metabolism, and behavior via transcriptional feedback loops. Because key circadian transcriptional activators and repressors are regulated by phosphorylation, we screened for phosphatases that alter activity rhythms when their expression was reduced. One such phosphatase, leukocyte-antigen-related (LAR), abolishes activity rhythms, but does not disrupt feedback loop function. Rather,Lardisrupts clock output by eliminating axonal processes from clock neurons that release pigment-dispersing factor (PDF) neuropeptide into the dorsal brain, but PDF expression persists in their cell bodies and remaining projections. In contrast to flies that lack PDF, flies that lackLaranticipate lights-on and lights-off transitions normally, which suggests that the remaining PDF expression mediates activity during light/dark cycles.
authors.affiliation: Department of Biology and Center for Biological Clocks Research, Texas A&M University, College Station, Texas 77845-3258.
authors.forename: Parul
authors.initials: P
authors.lastname: Agrawal
authors.affiliation: Department of Biology and Center for Biological Clocks Research, Texas A&M University, College Station, Texas 77845-3258 phardin@bio.tamu.edu.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Hardin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001683:Biological Clocks / Q000235:genetics; D001921:Brain / Q000378:metabolism; D002940:Circadian Rhythm / Q000235:genetics*; D016373:DNA, Antisense / Q000494:pharmacology; D003624:Darkness; D004330:Drosophila / Q000502:physiology*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D007814:Larva; D008297:Male; D009043:Motor Activity / Q000235:genetics; D009154:Mutation / Q000235:genetics; D009474:Neurons / Q000502:physiology*; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference / Q000502:physiology; D054557:Receptor-Like Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*
chemical_list: D016373:DNA, Antisense; D029721:Drosophila Proteins; D009479:Neuropeptides; C113883:pdf protein, Drosophila; C096445:tim protein, Drosophila; C117978:Lar protein, Drosophila; D054557:Receptor-Like Protein Tyrosine Phosphatases
keywords: circadian rhythms; clock genes; locomotor activity; neuropeptides; pacemaker neurons; phosphatase