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Kazuhiro Shimomura,
Vivek Kumar,
Nobuya Koike,
Tae-Kyung Kim,
Jason Chong, Ethan D Buhr,
Andrew R Whiteley,
Sharon S Low,
Chiaki Omura,
Deborah Fenner, [......],
Marc Richards,
Seung-Hee Yoo,
Hee-Kyung Hong,
Martha H Vitaterna,
Joseph Bass,
Mathew T Pletcher,
Tim Wiltshire,
John Hogenesch,
Phillip L Lowrey,
Joseph S Takahashi
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ABSTRACT: Genetic and molecular approaches have been critical for elucidating the mechanism of the mammalian circadian clock. Here, we demonstrate that the ClockΔ19 mutant behavioral phenotype is significantly modified by mouse strain genetic background. We map a suppressor of the ClockΔ19 mutation to a ∼900 kb interval on mouse chromosome 1 and identify the transcription factor, Usf1, as the responsible gene. A SNP in the promoter of Usf1 causes elevation of its transcript and protein in strains that suppress the Clock mutant phenotype. USF1 competes with the CLOCK:BMAL1 complex for binding to E-box sites in target genes. Saturation binding experiments demonstrate reduced affinity of the CLOCKΔ19:BMAL1 complex for E-box sites, thereby permitting increased USF1 occupancy on a genome-wide basis. We propose that USF1 is an important modulator of molecular and behavioral circadian rhythms in mammals. DOI:http://dx.doi.org/10.7554/eLife.00426.001.
eLife. 01/2013; 2:e00426.
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ABSTRACT: The amplitude of a circadian oscillator influences its response to a phase-resetting stimulus. The suprachiasmatic nucleus (SCN) is unique among circadian clocks in mammals in that the network connections among its neurons confer robustness both in amplitude and in resilience to perturbations. With reduced coupling among SCN neurons, the SCN becomes more susceptible to external phase-shifting stimuli. Thus, stimuli of the same strength will elicit different responses from the same tissue under different states of internal coupling. In his letter, Ruby (2011 [this issue]) discusses potential causes for discrepancies in studies that report dissimilar responses of the SCN to temperature changes. Here, we propose that the differences are likely due to a species difference and a difference in oscillator amplitude. These differences more likely result from inherent differences between mice and rats than from experimental procedures.
Journal of Biological Rhythms 08/2011; 26(4):371-3. · 2.93 Impact Factor
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ABSTRACT: Environmental temperature cycles are a universal entraining cue for all circadian systems at the organismal level with the exception of homeothermic vertebrates. We report here that resistance to temperature entrainment is a property of the suprachiasmatic nucleus (SCN) network and is not a cell-autonomous property of mammalian clocks. This differential sensitivity to temperature allows the SCN to drive circadian rhythms in body temperature, which can then act as a universal cue for the entrainment of cell-autonomous oscillators throughout the body. Pharmacological experiments show that network interactions in the SCN are required for temperature resistance and that the heat shock pathway is integral to temperature resetting and temperature compensation in mammalian cells. These results suggest that the evolutionarily ancient temperature resetting response can be used in homeothermic animals to enhance internal circadian synchronization.
Science 10/2010; 330(6002):379-85. · 31.20 Impact Factor
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Biliana Marcheva,
Kathryn Moynihan Ramsey, Ethan D Buhr,
Yumiko Kobayashi,
Hong Su,
Caroline H Ko,
Ganka Ivanova,
Chiaki Omura,
Shelley Mo,
Martha H Vitaterna,
James P Lopez,
Louis H Philipson,
Christopher A Bradfield,
Seth D Crosby,
Lellean JeBailey,
Xiaozhong Wang,
Joseph S Takahashi,
Joseph Bass
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ABSTRACT: The molecular clock maintains energy constancy by producing circadian oscillations of rate-limiting enzymes involved in tissue metabolism across the day and night. During periods of feeding, pancreatic islets secrete insulin to maintain glucose homeostasis, and although rhythmic control of insulin release is recognized to be dysregulated in humans with diabetes, it is not known how the circadian clock may affect this process. Here we show that pancreatic islets possess self-sustained circadian gene and protein oscillations of the transcription factors CLOCK and BMAL1. The phase of oscillation of islet genes involved in growth, glucose metabolism and insulin signalling is delayed in circadian mutant mice, and both Clock and Bmal1 (also called Arntl) mutants show impaired glucose tolerance, reduced insulin secretion and defects in size and proliferation of pancreatic islets that worsen with age. Clock disruption leads to transcriptome-wide alterations in the expression of islet genes involved in growth, survival and synaptic vesicle assembly. Notably, conditional ablation of the pancreatic clock causes diabetes mellitus due to defective beta-cell function at the very latest stage of stimulus-secretion coupling. These results demonstrate a role for the beta-cell clock in coordinating insulin secretion with the sleep-wake cycle, and reveal that ablation of the pancreatic clock can trigger the onset of diabetes mellitus.
Nature 07/2010; 466(7306):627-31. · 36.28 Impact Factor
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Kazuhiro Shimomura,
Phillip L Lowrey,
Martha Hotz Vitaterna, Ethan D Buhr,
Vivek Kumar,
Peter Hanna,
Chiaki Omura,
Mariko Izumo,
Sharon S Low,
R Keith Barrett,
Silvia I LaRue,
Carla B Green,
Joseph S Takahashi
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ABSTRACT: Most laboratory mouse strains including C57BL/6J do not produce detectable levels of pineal melatonin owing to deficits in enzymatic activity of arylalkylamine N-acetyltransferase (AANAT) and N-acetylserotonin O-methyl transferase (ASMT), two enzymes necessary for melatonin biosynthesis. Here we report that alleles segregating at these two loci in C3H/HeJ mice, an inbred strain producing melatonin, suppress the circadian period-lengthening effect of the Clock mutation. Through a functional mapping approach, we localize mouse Asmt to chromosome X and show that it, and the Aanat locus on chromosome 11, are significantly associated with pineal melatonin levels. Treatment of suprachiasmatic nucleus (SCN) explant cultures from Period2(Luciferase) (Per2(Luc)) Clock/+ reporter mice with melatonin, or the melatonin agonist, ramelteon, phenocopies the genetic suppression of the Clock mutant phenotype observed in living animals. These results demonstrate that melatonin suppresses the Clock/+ mutant phenotype and interacts with Clock to affect the mammalian circadian system.
Proceedings of the National Academy of Sciences 05/2010; 107(18):8399-403. · 9.68 Impact Factor
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PLoS Biology 01/2010; 8(10). · 11.45 Impact Factor
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ABSTRACT: Bmal1 is an essential transcriptional activator within the mammalian circadian clock. We report here that the suprachiasmatic nucleus (SCN) of Bmal1-null mutant mice, unexpectedly, generates stochastic oscillations with periods that overlap the circadian range. Dissociated SCN neurons expressed fluctuating levels of PER2 detected by bioluminescence imaging but could not generate circadian oscillations intrinsically. Inhibition of intercellular communication or cyclic-AMP signaling in SCN slices, which provide a positive feed-forward signal to drive the intracellular negative feedback loop, abolished the stochastic oscillations. Propagation of this feed-forward signal between SCN neurons then promotes quasi-circadian oscillations that arise as an emergent property of the SCN network. Experimental analysis and mathematical modeling argue that both intercellular coupling and molecular noise are required for the stochastic rhythms, providing a novel biological example of noise-induced oscillations. The emergence of stochastic circadian oscillations from the SCN network in the absence of cell-autonomous circadian oscillatory function highlights a previously unrecognized level of circadian organization.
PLoS Biology 01/2010; 8(10):e1000513. · 11.45 Impact Factor
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Kathryn Moynihan Ramsey,
Jun Yoshino,
Cynthia S Brace,
Dana Abrassart,
Yumiko Kobayashi,
Biliana Marcheva,
Hee-Kyung Hong,
Jason L Chong, Ethan D Buhr,
Choogon Lee,
Joseph S Takahashi,
Shin-Ichiro Imai,
Joseph Bass
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ABSTRACT: The circadian clock is encoded by a transcription-translation feedback loop that synchronizes behavior and metabolism with the light-dark cycle. Here we report that both the rate-limiting enzyme in mammalian nicotinamide adenine dinucleotide (NAD+) biosynthesis, nicotinamide phosphoribosyltransferase (NAMPT), and levels of NAD+ display circadian oscillations that are regulated by the core clock machinery in mice. Inhibition of NAMPT promotes oscillation of the clock gene Per2 by releasing CLOCK:BMAL1 from suppression by SIRT1. In turn, the circadian transcription factor CLOCK binds to and up-regulates Nampt, thus completing a feedback loop involving NAMPT/NAD+ and SIRT1/CLOCK:BMAL1.
Science 04/2009; 324(5927):651-4. · 31.20 Impact Factor
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Andrew C Liu,
David K Welsh,
Caroline H Ko,
Hien G Tran,
Eric E Zhang,
Aaron A Priest, Ethan D Buhr,
Oded Singer,
Kirsten Meeker,
Inder M Verma,
Francis J Doyle,
Joseph S Takahashi,
Steve A Kay
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ABSTRACT: Molecular mechanisms of the mammalian circadian clock have been studied primarily by genetic perturbation and behavioral analysis. Here, we used bioluminescence imaging to monitor Per2 gene expression in tissues and cells from clock mutant mice. We discovered that Per1 and Cry1 are required for sustained rhythms in peripheral tissues and cells, and in neurons dissociated from the suprachiasmatic nuclei (SCN). Per2 is also required for sustained rhythms, whereas Cry2 and Per3 deficiencies cause only period length defects. However, oscillator network interactions in the SCN can compensate for Per1 or Cry1 deficiency, preserving sustained rhythmicity in mutant SCN slices and behavior. Thus, behavior does not necessarily reflect cell-autonomous clock phenotypes. Our studies reveal previously unappreciated requirements for Per1, Per2, and Cry1 in sustaining cellular circadian rhythmicity and demonstrate that SCN intercellular coupling is essential not only to synchronize component cellular oscillators but also for robustness against genetic perturbations.
Cell 06/2007; 129(3):605-16. · 32.40 Impact Factor
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ABSTRACT: The mouse Clock gene encodes a basic helix-loop-helix-PAS transcription factor, CLOCK, that acts in concert with BMAL1 to form the positive elements of the circadian clock mechanism in mammals. The original Clock mutant allele is a dominant negative (antimorphic) mutation that deletes exon 19 and causes an internal deletion of 51 aa in the C-terminal activation domain of the CLOCK protein. Here we report that heterozygous Clock/+ mice exhibit high-amplitude phase-resetting responses to 6-h light pulses (Type 0 resetting) as compared with wild-type mice that have low amplitude (Type 1) phase resetting. The magnitude and time course of acute light induction in the suprachiasmatic nuclei of the only known light-induced core clock genes, Per1 and Per2, are not affected by the Clock/+ mutation. However, the amplitude of the circadian rhythms of Per gene expression are significantly reduced in Clock homozygous and heterozygous mutants. Rhythms of PER2::LUCIFERASE expression in suprachiasmatic nuclei explant cultures also are reduced in amplitude in Clock heterozygotes. The phase-response curves to changes in culture medium are Type 0 in Clock heterozygotes, but Type 1 in wild types, similar to that seen for light in vivo. The increased efficacy of resetting stimuli and decreased PER expression amplitude can be explained in a unified manner by a model in which the Clock mutation reduces circadian pacemaker amplitude in the suprachiasmatic nuclei.
Proceedings of the National Academy of Sciences 07/2006; 103(24):9327-32. · 9.68 Impact Factor
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ABSTRACT: The mouse Period2 (mPer2) locus is an essential negative-feedback element of the mammalian circadian-clock mechanism. Recent work has shown that mPer2 circadian gene expression persists in both central and peripheral tissues. Here, we analyze the mouse mPer2 promoter and identify a circadian enhancer (E2) with a noncanonical 5'-CACGTT-3' E-box located 20 bp upstream of the mPer2 transcription start site. The E2 enhancer accounts for most circadian transcriptional drive of the mPer2 locus by CLOCK:BMAL1, is a major site of DNaseI hypersensitivity in this region, and is constitutively bound by a transcriptional complex containing the CLOCK protein. Importantly, the E2 enhancer is sufficient to drive self-sustained circadian rhythms of luciferase activity in central and peripheral tissues from mPer2-E2::Luciferase transgenic mice with tissue-specific phase and period characteristics. Last, genetic analysis with mutations in Clock and Bmal1 shows that the E2 enhancer is a target of CLOCK and BMAL1 in vivo.
Proceedings of the National Academy of Sciences 03/2005; 102(7):2608-13. · 9.68 Impact Factor
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Seung-Hee Yoo,
Shin Yamazaki,
Phillip L Lowrey,
Kazuhiro Shimomura,
Caroline H Ko, Ethan D Buhr,
Sandra M Siepka,
Hee-Kyung Hong,
Won Jun Oh,
Ook Joon Yoo,
Michael Menaker,
Joseph S Takahashi
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ABSTRACT: Mammalian circadian rhythms are regulated by the suprachiasmatic nucleus (SCN), and current dogma holds that the SCN is required for the expression of circadian rhythms in peripheral tissues. Using a PERIOD2::LUCIFERASE fusion protein as a real-time reporter of circadian dynamics in mice, we report that, contrary to previous work, peripheral tissues are capable of self-sustained circadian oscillations for >20 cycles in isolation. In addition, peripheral organs expressed tissue-specific differences in circadian period and phase. Surprisingly, lesions of the SCN in mPer2(Luciferase) knockin mice did not abolish circadian rhythms in peripheral tissues, but instead caused phase desynchrony among the tissues of individual animals and from animal to animal. These results demonstrate that peripheral tissues express self-sustained, rather than damped, circadian oscillations and suggest the existence of organ-specific synchronizers of circadian rhythms at the cell and tissue level.
Proceedings of the National Academy of Sciences 05/2004; 101(15):5339-46. · 9.68 Impact Factor
