Quantitative live imaging of Venus::BMAL1 in a mouse model reveals complex dynamics of the master circadian clock regulator.

Autor: Yang N; Wellcome Centre for Cell Matrix Research, University of Manchester, Manchester, United Kingdom.; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Smyllie NJ; Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, United Kingdom., Morris H; Wellcome Centre for Cell Matrix Research, University of Manchester, Manchester, United Kingdom.; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Gonçalves CF; Wellcome Centre for Cell Matrix Research, University of Manchester, Manchester, United Kingdom.; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Dudek M; Wellcome Centre for Cell Matrix Research, University of Manchester, Manchester, United Kingdom.; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Pathiranage DRJ; Wellcome Centre for Cell Matrix Research, University of Manchester, Manchester, United Kingdom.; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Chesham JE; Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, United Kingdom., Adamson A; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Spiller DG; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Zindy E; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Bagnall J; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Humphreys N; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Hoyland J; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom.; NIHR Manchester Musculoskeletal Biomedical Research Centre, Manchester University NHS Foundation Trust, Manchester Academic Health Science Centre, Manchester, United Kingdom., Loudon ASI; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom., Hastings MH; Division of Neurobiology, Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, United Kingdom., Meng QJ; Wellcome Centre for Cell Matrix Research, University of Manchester, Manchester, United Kingdom.; Faculty of Biology, Medicine and Health, University of Manchester, Manchester, United Kingdom.
Jazyk: angličtina
Zdroj: PLoS genetics [PLoS Genet] 2020 Apr 30; Vol. 16 (4), pp. e1008729. Date of Electronic Publication: 2020 Apr 30 (Print Publication: 2020).
DOI: 10.1371/journal.pgen.1008729
Abstrakt: Evolutionarily conserved circadian clocks generate 24-hour rhythms in physiology and behaviour that adapt organisms to their daily and seasonal environments. In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus is the principal co-ordinator of the cell-autonomous clocks distributed across all major tissues. The importance of robust daily rhythms is highlighted by experimental and epidemiological associations between circadian disruption and human diseases. BMAL1 (a bHLH-PAS domain-containing transcription factor) is the master positive regulator within the transcriptional-translational feedback loops (TTFLs) that cell-autonomously define circadian time. It drives transcription of the negative regulators Period and Cryptochrome alongside numerous clock output genes, and thereby powers circadian time-keeping. Because deletion of Bmal1 alone is sufficient to eliminate circadian rhythms in cells and the whole animal it has been widely used as a model for molecular disruption of circadian rhythms, revealing essential, tissue-specific roles of BMAL1 in, for example, the brain, liver and the musculoskeletal system. Moreover, BMAL1 has clock-independent functions that influence ageing and protein translation. Despite the essential role of BMAL1 in circadian time-keeping, direct measures of its intra-cellular behaviour are still lacking. To fill this knowledge-gap, we used CRISPR Cas9 to generate a mouse expressing a knock-in fluorescent fusion of endogenous BMAL1 protein (Venus::BMAL1) for quantitative live imaging in physiological settings. The Bmal1Venus mouse model enabled us to visualise and quantify the daily behaviour of this core clock factor in central (SCN) and peripheral clocks, with single-cell resolution that revealed its circadian expression, anti-phasic to negative regulators, nuclear-cytoplasmic mobility and molecular abundance.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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