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1 Division of Molecular and Cellular Physiology, Department of Physiological Sciences, Lund University, Lund, Sweden
2 Department of Cardiology, Lund University, Lund, Sweden
3 Department of Cardiothoracic Surgery, Lund University, Lund, Sweden
4 School of Biomedical Sciences, University of Leeds, Leeds, United Kingdom
* To whom correspondence should be addressed. E-mail: Per.Hellstrand{at}mphy.lu.se.
Loss of the smooth muscle contractile phenotype is critical in atherosclerosis and in restenosis following angioplasty, but its early signals are incompletely understood. Here we explore the role of TRPC proteins, suggested to mediate store-operated Ca2+ entry (SOCE). Contractility of rat cerebral arteries in organ culture is preserved for several days, whereas SOCE is increased. Correlating with this, nifedipine-insensitive whole-cell current, activated by depletion of intracellular Ca2+ stores, was increased by 50% in cells isolated from arteries cultured for 3 days. TRPC1 and -6 mRNA were >5-fold increased in cells isolated after organ culture, whereas TRPC3 was decreased. Immunofluorescence staining and/or western blots of arteries and isolated cells showed up-regulation of TRPC1 and -6 proteins during organ culture. In intact arteries, TRPC4 expression correlated with the amount of endothelium present. Ca2+ addition following store depletion caused a contraction in cultured but not freshly dissected arteries. A polyclonal TRPC1 antibody directed against an extracellular epitope inhibited this contraction by ~50%. To investigate the basis of the TRPC up-regulation and assess its possible clinical significance, segments of human internal mammary artery were organ-cultured for 24 h and then exposed to balloon dilatation in vitro, followed by further culture for up to 48 h. Following dilatation, TRPC1 and -6 mRNA were progressively increased in comparison with undilated control segments. The results of this study indicate that vascular injury enhances plasticity in TRPC expression, that TRPC expression correlates with cellular Ca2+ handling, and that TRPC1 is a subunit of up-regulated SOC channels.
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