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NEUROHUMORAL CONTROL OF CARDIOVASCULAR FUNCTION

Vascular circadian rhythms in a mouse vascular smooth muscle cell line (Movas-1)

¹Departments of Physiology, ²Medicine, and Ob/Gyn, University of Toronto, Medical Sciences Building; ³Heart and Stroke Richard Lewar Centre for Cardiovascular Excellence, and Toronto General Hospital Research Institute; and ⁴Centre for Biological Timing and Cognition, Department of Psychology, University of Toronto, Toronto, Ontario, Canada

Submitted 4 July 2008 ; accepted in final form 26 August 2008

The circadian system in mammals is a hierarchy of oscillators throughout the organism that are coordinated by the circadian clock in the hypothalamic suprachiasmatic nucleus. Peripheral clocks act to integrate time-of-day information from neural or hormonal signals, regulating gene expression, and, subsequently, organ physiology. However, the mechanisms by which the central clock communicates with peripheral oscillators are not understood and are likely tissue specific. In this study, we establish a mouse vascular cell model suitable for investigations of these mechanisms at a molecular level. Using the immortalized vascular smooth muscle cell line Movas-1, we determined that these cells express the circadian clock machinery with robust rhythms in mRNA expression over a 36-h period after serum shock synchronization. Furthermore, norepinephrine and forskolin were able to synchronize circadian rhythms in bmal1. With synchronization, we observed cycling of specific genes, including the tissue inhibitor of metalloproteinase 1 and 3 (timp1, timp3), collagen 3a1 (col3a1), transgelin 1 (sm22), and calponin 1 (cnn1). Diurnal expression of these genes was also found in vivo in mouse aortic tissue, using microarray and real-time RT-PCR analysis. Both of these revealed ultradian rhythms in genes similar to the cycling observed in Movas-1 in vitro. These findings highlight the cyclical nature of structurally important genes in the vasculature that is similar both in vivo and in vitro. This study establishes the Movas-1 cells as a novel cell model from which to further investigate the molecular mechanisms of clock regulation in the vasculature.

real-time RT-PCR; ultradian; diurnal; microarray

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