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Vascular smooth muscle cell calcification and SLC20 inorganic phosphate transporters: effects of PDGF, TNF-α, and Pi

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Abstract

Pi transport by vascular smooth muscle cells (VSMC) has been proposed to play an important role in the pathogenesis of vascular calcification. In this study, we have determined the correlation between calcification induced by Pi, platelet-derived growth factor (PDGF)-BB, and tumor necrosis factor-α and Pi transport activity in primary cultures of rat aortic VSMC. These agents induced calcification and increased the expression of Cbfa1, Msx2, and Bmp2 osteogene messenger RNA in rat aortic VSMC, while Pi transport rate was not modified per milligram of protein. Only PDGF increased Pi transport when it was expressed per unit of DNA, as PDGF also increased total cell protein by 100%, while DNA content and number of cells were not modified. PDGF increased the expression of the Pi transporter, Pit-1, but membrane protein biotinylation showed that Pit-1 abundance was not modified in the cell surface. Immunofluorescence revealed that, under basal conditions, Pit-1 is only slightly expressed at the cell membrane, but strongly expressed inside the cell. The intracellular signal colocalizes with endoplasmic reticulum (ER) markers, and PDGF increases Pit-1 expression in the ER but not the cell membrane. In conclusion, Pi transport across the plasma membrane does not correlate directly with calcification, but the expression of Pit-1 in the ER opens new possibilities for the study of the pathogenesis of vascular calcification.

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Acknowledgments

This work was supported by grants BFU2006-06284/BFI from the Spanish Ministry of Education and Science to VS, 1 R01 DK066029 from the US National Institute of Health to ML, and predoctoral fellowship B086/2007 from the Government of Aragón (Spain) to RVB.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratory of Molecular Toxicology, Veterinary Faculty, University of Zaragoza, Calle Miguel Servet 177, 50013, Zaragoza, Spain

    Ricardo Villa-Bellosta & Victor Sorribas

  2. Department of Medicine, Division of Renal Diseases and Hypertension, University of Colorado Denver and Denver VAMC, 12700 East 19th Avenue, Research 2, Room 7002, Box C281, Aurora, CO, 80045, USA

    Moshe Levi & Victor Sorribas

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  1. Ricardo Villa-Bellosta
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  2. Moshe Levi
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Correspondence to Victor Sorribas.

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Villa-Bellosta, R., Levi, M. & Sorribas, V. Vascular smooth muscle cell calcification and SLC20 inorganic phosphate transporters: effects of PDGF, TNF-α, and Pi. Pflugers Arch - Eur J Physiol 458, 1151–1161 (2009). https://doi.org/10.1007/s00424-009-0688-5

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  • DOI: https://doi.org/10.1007/s00424-009-0688-5

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