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Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures

BACKGROUND: Hepatocytes, the parenchymal cells of the liver, express core clock genes, such as Period2 and Cryptochrome2, which are involved in the transcriptional/translational feedback loop of the circadian clock. Whether or not the liver is capable of sustaining rhythms independent of a central p...

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Autores principales: Guenthner, Casey J., Luitje, Martha E., Pyle, Lorna A., Molyneux, Penny C., Yu, Jimmy K., Li, Alexander S., Leise, Tanya L., Harrington, Mary E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3911982/
https://www.ncbi.nlm.nih.gov/pubmed/24498336
http://dx.doi.org/10.1371/journal.pone.0087573
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author Guenthner, Casey J.
Luitje, Martha E.
Pyle, Lorna A.
Molyneux, Penny C.
Yu, Jimmy K.
Li, Alexander S.
Leise, Tanya L.
Harrington, Mary E.
author_facet Guenthner, Casey J.
Luitje, Martha E.
Pyle, Lorna A.
Molyneux, Penny C.
Yu, Jimmy K.
Li, Alexander S.
Leise, Tanya L.
Harrington, Mary E.
author_sort Guenthner, Casey J.
collection PubMed
description BACKGROUND: Hepatocytes, the parenchymal cells of the liver, express core clock genes, such as Period2 and Cryptochrome2, which are involved in the transcriptional/translational feedback loop of the circadian clock. Whether or not the liver is capable of sustaining rhythms independent of a central pacemaker is controversial. Whether and how circadian information may be shared among cells in the liver in order to sustain oscillations is currently unknown. RESULTS: In this study we isolated primary hepatocytes from transgenic Per2(Luc) mice and used bioluminescence as a read-out of the state of the circadian clock. Hepatocytes cultured in a collagen gel sandwich configuration exhibited persistent circadian rhythms for several weeks. The amplitude of the rhythms damped, but medium changes consistently reset the phase and amplitude of the cultures. Cry2(−/−) Per2(Luc) cells oscillated robustly and expressed a longer period. Co-culturing with wildtype cells did not significantly shorten the period, indicating that coupling among hepatocytes is insufficient to synchronize cells with significantly differing periods. However, spatial patterns revealed by cellular imaging of wildtype cultures provided evidence of weak local coupling among the hepatocytes. CONCLUSIONS: Our results with primary hepatocyte cultures demonstrate that cultured hepatocytes are weakly coupled. While this coupling is not sufficient to sustain global synchrony, it does increase local synchrony, which may stabilize the circadian rhythms of peripheral oscillators, such as the liver, against noise in the entraining signals.
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spelling pubmed-39119822014-02-04 Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures Guenthner, Casey J. Luitje, Martha E. Pyle, Lorna A. Molyneux, Penny C. Yu, Jimmy K. Li, Alexander S. Leise, Tanya L. Harrington, Mary E. PLoS One Research Article BACKGROUND: Hepatocytes, the parenchymal cells of the liver, express core clock genes, such as Period2 and Cryptochrome2, which are involved in the transcriptional/translational feedback loop of the circadian clock. Whether or not the liver is capable of sustaining rhythms independent of a central pacemaker is controversial. Whether and how circadian information may be shared among cells in the liver in order to sustain oscillations is currently unknown. RESULTS: In this study we isolated primary hepatocytes from transgenic Per2(Luc) mice and used bioluminescence as a read-out of the state of the circadian clock. Hepatocytes cultured in a collagen gel sandwich configuration exhibited persistent circadian rhythms for several weeks. The amplitude of the rhythms damped, but medium changes consistently reset the phase and amplitude of the cultures. Cry2(−/−) Per2(Luc) cells oscillated robustly and expressed a longer period. Co-culturing with wildtype cells did not significantly shorten the period, indicating that coupling among hepatocytes is insufficient to synchronize cells with significantly differing periods. However, spatial patterns revealed by cellular imaging of wildtype cultures provided evidence of weak local coupling among the hepatocytes. CONCLUSIONS: Our results with primary hepatocyte cultures demonstrate that cultured hepatocytes are weakly coupled. While this coupling is not sufficient to sustain global synchrony, it does increase local synchrony, which may stabilize the circadian rhythms of peripheral oscillators, such as the liver, against noise in the entraining signals. Public Library of Science 2014-02-03 /pmc/articles/PMC3911982/ /pubmed/24498336 http://dx.doi.org/10.1371/journal.pone.0087573 Text en © 2014 Guenthner et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Guenthner, Casey J.
Luitje, Martha E.
Pyle, Lorna A.
Molyneux, Penny C.
Yu, Jimmy K.
Li, Alexander S.
Leise, Tanya L.
Harrington, Mary E.
Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures
title Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures
title_full Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures
title_fullStr Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures
title_full_unstemmed Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures
title_short Circadian Rhythms of PER2::LUC in Individual Primary Mouse Hepatocytes and Cultures
title_sort circadian rhythms of per2::luc in individual primary mouse hepatocytes and cultures
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3911982/
https://www.ncbi.nlm.nih.gov/pubmed/24498336
http://dx.doi.org/10.1371/journal.pone.0087573
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