Electrical Silencing of Drosophila Pacemaker Neurons Stops the Free-Running Circadian Clock

Electrical silencing of Drosophila circadian pacemaker neurons through targeted expression of K+ channels causes severe deficits in free-running circadian locomotor rhythmicity in complete darkness. Pacemaker electrical silencing also stops the free-running oscillation of PERIOD (PER) and TIMELESS (TIM) proteins that constitutes the core of the cell-autonomous molecular clock. In contrast, electrical silencing fails to abolish PER and TIM oscillation in light-dark cycles, although it does impair rhythmic behavior. On the basis of these findings, we propose that electrical activity is an essential element of the free-running molecular clock of pacemaker neurons along with the transcription factors and regulatory enzymes that have been previously identified as required for clock function.

[1]  J. C. Hall,et al.  Disruption of synaptic transmission or clock-gene-product oscillations in circadian pacemaker cells of Drosophila cause abnormal behavioral rhythms. , 2000, Journal of neurobiology.

[2]  S. Goldstein,et al.  ORK1, a potassium-selective leak channel with two pore domains cloned from Drosophila melanogaster by expression in Saccharomyces cerevisiae. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[3]  I. Levitan,et al.  It Is Calmodulin After All! Mediator of the Calcium Modulation of Multiple Ion Channels , 1999, Neuron.

[4]  S. Kay,et al.  Time zones: a comparative genetics of circadian clocks , 2001, Nature Reviews Genetics.

[5]  B. Gold The pathophysiology of proximal neurofilamentous giant axonal swellings: implications for the pathogenesis of amyotrophic lateral sclerosis. , 1987, Toxicology.

[6]  S. Reppert,et al.  Comparing Clockworks: Mouse versus Fly , 2000, Journal of biological rhythms.

[7]  T. Préat,et al.  Defining the role of Drosophila lateral neurons in the control of circadian rhythms in motor activity and eclosion by targeted genetic ablation and PERIOD protein overexpression , 2001, The European journal of neuroscience.

[8]  Edmund M Talley,et al.  TASK-1, a Two–Pore Domain K+ Channel, Is Modulated by Multiple Neurotransmitters in Motoneurons , 2000, Neuron.

[9]  G. Davis,et al.  Homeostatic Control of Presynaptic Release Is Triggered by Postsynaptic Membrane Depolarization , 2001, Neuron.

[10]  Jeffrey C. Hall,et al.  Drosophila CRY Is a Deep Brain Circadian Photoreceptor , 2000, Neuron.

[11]  P. Sokolove,et al.  The chi square periodogram: its utility for analysis of circadian rhythms. , 1978, Journal of theoretical biology.

[12]  Hong Zhou,et al.  The Drosophila dCREB2 Gene Affects the Circadian Clock , 1999, Neuron.

[13]  J. C. Hall,et al.  Differential regulation of circadian pacemaker output by separate clock genes in Drosophila. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. W. Young,et al.  double-time Is a Novel Drosophila Clock Gene that Regulates PERIOD Protein Accumulation , 1998, Cell.

[15]  Leonard K. Kaczmarek,et al.  Targeted Attenuation of Electrical Activity in Drosophila Using a Genetically Modified K+ Channel , 2001, Neuron.

[16]  K R Svoboda,et al.  Activity regulates programmed cell death of zebrafish Rohon-Beard neurons. , 2001, Development.

[17]  Michael J. McDonald,et al.  Microarray Analysis and Organization of Circadian Gene Expression in Drosophila , 2001, Cell.

[18]  I. Levitan,et al.  Association of Src Tyrosine Kinase with a Human Potassium Channel Mediated by SH3 Domain , 1996, Science.

[19]  M. Barish Intracellular calcium regulation of channel and receptor expression in the plasmalemma: potential sites of sensitivity along the pathways linking transcription, translation, and insertion. , 1998, Journal of neurobiology.

[20]  Jeffrey C. Hall,et al.  The Circadian Clock of Fruit Flies Is Blind after Elimination of All Known Photoreceptors , 2001, Neuron.

[21]  J. C. Hall,et al.  Involvement of the period Gene in Developmental Time-Memory: Effect of the perShort Mutation on Phase Shifts Induced by Light Pulses Delivered to Drosophila Larvae , 2000, Journal of biological rhythms.

[22]  C. Helfrich-Förster The period clock gene is expressed in central nervous system neurons which also produce a neuropeptide that reveals the projections of circadian pacemaker cells within the brain of Drosophila melanogaster. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[23]  M. W. Young The molecular control of circadian behavioral rhythms and their entrainment in Drosophila. , 1998, Annual review of biochemistry.

[24]  Markus Meister,et al.  Individual neurons dissociated from rat suprachiasmatic nucleus express independently phased circadian firing rhythms , 1995, Neuron.

[25]  Jeffrey C. Hall,et al.  A pdf neuropeptide gene mutation and ablation of PDF neurons each cause severe abnormalities of behavioral circadian rhythms in Drosophila. , 2000, Cell.

[26]  J. C. Hall,et al.  Spatial and Temporal Expression of the period andtimeless Genes in the Developing Nervous System ofDrosophila: Newly Identified Pacemaker Candidates and Novel Features of Clock Gene Product Cycling , 1997, The Journal of Neuroscience.

[27]  B. Robertson,et al.  A functional role for the two-pore domain potassium channel TASK-1 in cerebellar granule neurons. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[28]  G. Block,et al.  A Delayed Rectifier Current Is Modulated by the Circadian Pacemaker in Bulla , 1999, Journal of biological rhythms.

[29]  A. West,et al.  Calcium regulation of neuronal gene expression , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. Mains,et al.  Inducible Genetic Suppression of Neuronal Excitability , 1999, The Journal of Neuroscience.

[31]  Michael Bate,et al.  Altered Electrical Properties in DrosophilaNeurons Developing without Synaptic Transmission , 2001, The Journal of Neuroscience.

[32]  S. Carpenter,et al.  Neurofibrillary axonal swellings and amyotrophic lateral sclerosis , 1984, Journal of the Neurological Sciences.

[33]  H. Lester,et al.  ROMK1 (Kir1.1) causes apoptosis and chronic silencing of hippocampal neurons. , 2000, Journal of neurophysiology.

[34]  C. Colwell Circadian modulation of calcium levels in cells in the suprachiasmatic nucleus , 2000, The European journal of neuroscience.

[35]  W. Catterall Structure and regulation of voltage-gated Ca2+ channels. , 2000, Annual review of cell and developmental biology.

[36]  A. Scheibel,et al.  Degeneration and regeneration of the nervous system , 1960 .

[37]  M. Nitabach,et al.  A mechanism for combinatorial regulation of electrical activity: Potassium channel subunits capable of functioning as Src homology 3-dependent adaptors. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[38]  C. Goodman,et al.  Ectopic and increased expression of fasciclin II alters motoneuron growth cone guidance , 1994, Neuron.

[39]  W. Schwartz Suprachiasmatic nucleus , 2002, Current Biology.

[40]  A. Mccarthy Development , 1996, Current Opinion in Neurobiology.

[41]  S. Goldstein,et al.  Opening and Closing of KcnkØ Potassium Leak Channels Is Tightly Regulated , 2000, The Journal of general physiology.

[42]  Adam Claridge‐Chang,et al.  Circadian Regulation of Gene Expression Systems in the Drosophila Head , 2001, Neuron.

[43]  K. Yao,et al.  Neural Specificity of elav Expression: Defining a Drosophila Promoter for Directing Expression to the Nervous System , 1994, Journal of neurochemistry.

[44]  G. Block,et al.  Circadian rhythm in membrane conductance expressed in isolated neurons. , 1993, Science.

[45]  J. Ruppersberg,et al.  Intracellular regulation of inward rectifier K+ channels , 2000, Pflügers Archiv.

[46]  Jeffrey C. Hall,et al.  CRY, a Drosophila Clock and Light-Regulated Cryptochrome, Is a Major Contributor to Circadian Rhythm Resetting and Photosensitivity , 1998, Cell.

[47]  A. Sehgal,et al.  Molecular components of the circadian system in Drosophila. , 2001, Annual review of physiology.

[48]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[49]  Jeffrey C. Hall,et al.  The cryb Mutation Identifies Cryptochrome as a Circadian Photoreceptor in Drosophila , 1998, Cell.

[50]  C. Colwell NMDA‐evoked calcium transients and currents in the suprachiasmatic nucleus: gating by the circadian system , 2001, The European journal of neuroscience.