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Modification of the Per2 clock gene in mPer2Luc reporter mice significantly alters circadian function. Behavioral period in constant dark is lengthened, and dissociates into two distinct components in constant light. Rhythms exhibit increased bimodality, enhanced phase resetting to light pulses, and altered entrainment to scheduled feeding. Mechanistic mathematical modelling predicts that enhanced protein interactions with the modified mPER2 C-terminus, combined with differential clock regulation among SCN subregions, can account for effects on circadian behavior via increased Per2 transcript and protein stability. PER2::LUC produces greater suppression of CLOCK:BMAL1 E-box activity than PER2. mPer2Luc carries a 72 bp deletion in exon 23 of Per2, and retains a neomycin resistance cassette that affects rhythm amplitude but not period. The results show that mPer2Luc acts as a circadian clock mutation illustrating a need for detailed assessment of potential impacts of c-terminal tags in genetically modified animal models. Author summary: Engineered genes that are modified to express bioluminescent signals can be used to monitor cellular processes by reflecting the actual state of the process in real time. For circadian rhythm studies, a widely used reporter model (mPer2Luc) expresses firefly luciferase under the control of the clock gene Period 2 (Per2), encoding a fusion protein (PER2::LUCIFERASE) which produces rhythms of bioluminescence. We present evidence that the engineered protein produces extensive changes in iconic circadian behavioral patterns in mice. The evidence comes from analysis of long-term recordings of circadian behavior in mPer2Luc mice in constant darkness and constant light, together with responsiveness to light pulses and entrainment to restricted food availability. By integrating a well-established mathematical model of circadian rhythm generation in the suprachiasmatic nuclei with the known functional heterogeneity within the nucleus, we recapitulated the observed locomotor behavioral patterns in WT and mPer2Luc mice. Functional alterations to the stability of PER2 protein predicted from the model were substantiated by increased repression strength of PER2::LUC over WT PER2 on CLOCK:BMAL1 transcriptional activation. Therefore, mPer2Luc is a clock mutation that provides new insights into the functional neuroanatomy of the mammalian circadian system and the critical importance of Per2 regulation in rhythm generation. [ABSTRACT FROM AUTHOR]

Circadian rhythmicity governs a remarkable array of fundamental biological functions and is mediated by cyclical transcriptomic and proteomic activities. Epigenetic factors are also involved in this circadian machinery; however, despite extensive efforts, detection and characterization of circadian cytosine modifications at the nucleotide level have remained elusive. In this study, we report that a large proportion of epigenetically variable cytosines show a circadian pattern in their modification status in mice. Importantly, the cytosines with circadian epigenetic oscillations significantly overlap with the cytosines exhibiting age-related changes in their modification status. Our findings suggest that evolutionary advantageous processes such as circadian rhythmicity can also contribute to an organism’s deterioration. [ABSTRACT FROM AUTHOR]

The above article from British Journal of Pharmacology, published online as an Accepted Article on 19 August 2019 in Wiley Online Library (wileyonlinelibrary.com), has been withdrawn by agreement between the authors, the journal Editor-in-Chief Professor Amrita Ahluwalia, and John Wiley & Sons Limited. The withdrawal has been agreed owing to new findings that necessitate re-interpretation of the results. [ABSTRACT FROM AUTHOR]

Circadian rhythmic expression of conditioned place avoidance (CPA) was produced in Syrian hamsters homozygous for the circadian short period mutation, tau. In constant dim red light neither the 20 h endogenous period, nor a 20 h place conditioning schedule eliminated the 24 h modulation of CPA behavior described previously for wild type (wt) hamsters and other species. Tau mutants exhibited a 20 h rhythm superimposed on the 24 h modulation. The 20 h component was removed selectively with lesions of the suprachiasmatic nucleus. Wt animals conditioned on a 20 h schedule did not produce a 20 h rhythm, but still expressed the 24 h modulation. The results show that the context entrainable oscillator (CEO) underlying memory for the timing of an unconditioned stimulus, retains a period of about 24 h regardless of clock gene background (tau mutation) and/or the conditioning schedule (24 vs 20 h). Therefore the CEO responsible for time memory is distinct from the biological clock controlling activity; the underlying circadian molecular mechanisms may differ from the ubiquitous transcription-translation feedback oscillator; and time memory itself is not classically conditioned. [ABSTRACT FROM AUTHOR]