Bioluminescence imaging of individual fibroblasts reveals persistent, independently phased circadian rhythms of clock gene expression

David K. Welsh, Seung Hee Yoo, Andrew C. Liu, Joseph S. Takahashi, Steve A. Kay

Research output: Contribution to journalArticle

449 Citations (Scopus)

Abstract

Circadian (ca. 24 hr) oscillations in expression of mammalian "clock genes" are found not only in the suprachiasmatic nucleus (SCN), the central circadian pacemaker, but also in peripheral tissues [1]. Under constant conditions in vitro, however, rhythms of peripheral tissue explants [2] or immortalized cells [3] damp partially or completely. It is unknown whether this reflects an inability of peripheral cells to sustain rhythms, as SCN neurons can, or a loss of synchrony among cells. Using bioluminescence imaging of Rat-1 fibroblasts transfected with a Bmal1::luc plasmid and primary fibroblasts dissociated from mPer2Luciferase-SV40 knockin mice, we monitored single-cell circadian rhythms of clock gene expression for 1-2 weeks. We found that single fibroblasts can oscillate robustly and independently with undiminished amplitude and diverse circadian periods. Cells were partially synchronized by medium changes at the start of an experiment, but due to different intrinsic periods, their phases became randomly distributed after several days. Closely spaced cells in the same culture did not have similar phases, implying a lack of functional coupling among cells. Thus, like SCN neurons, single fibroblasts can function as independent circadian oscillators; however, lack of oscillator coupling in dissociated cell cultures leads to a loss of synchrony among individual cells and damping of the ensemble rhythm at the population level.

Original languageEnglish (US)
Pages (from-to)2289-2295
Number of pages7
JournalCurrent Biology
Volume14
Issue number24
DOIs
StatePublished - Dec 29 2004

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Bioluminescence
Circadian Clocks
bioluminescence
Fibroblasts
Circadian Rhythm
Gene expression
circadian rhythm
fibroblasts
Clocks
image analysis
Gene Expression
Imaging techniques
gene expression
Suprachiasmatic Nucleus
Neurons
cells
Tissue
Pacemakers
Cell culture
cell culture

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Bioluminescence imaging of individual fibroblasts reveals persistent, independently phased circadian rhythms of clock gene expression. / Welsh, David K.; Yoo, Seung Hee; Liu, Andrew C.; Takahashi, Joseph S.; Kay, Steve A.

In: Current Biology, Vol. 14, No. 24, 29.12.2004, p. 2289-2295.

Research output: Contribution to journalArticle

Welsh, David K. ; Yoo, Seung Hee ; Liu, Andrew C. ; Takahashi, Joseph S. ; Kay, Steve A. / Bioluminescence imaging of individual fibroblasts reveals persistent, independently phased circadian rhythms of clock gene expression. In: Current Biology. 2004 ; Vol. 14, No. 24. pp. 2289-2295.
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