Evidence that TRPC3 is a molecular component of the 1alpha,25(OH)2D3-activated capacitative calcium entry (CCE) in muscle and osteoblast cells

The Journal of Steroid Biochemistry and Molecular Biology
Graciela SantillánRicardo Boland

Abstract

In chick skeletal muscle and in rat osteoblast-like cells (ROS 17/2.8), 1alpha,25-dihydroxy-Vitamin-D(3) [1alpha,25(OH)(2)D(3)] stimulates release of Ca(2+) from inner stores and extracellular cation influx through both voltage-dependent and capacitative Ca(2+) entry (CCE) channels. We investigated the involvement of TRPC proteins in CCE induced by 1alpha,25(OH)(2)D(3). Two fragments were amplified by RT-PCR, exhibiting >85% sequence homology with human TRPC3. Northern and Western blots employing TRPC3-probes and anti-TRPC3 antibodies, respectively, confirmed endogenous expression of a TRPC3-like protein. Both cell types transfected with anti-TRPC3 antisense oligodeoxynucleotides showed reduced CCE and Mn(2+) entry induced by either thapsigargin or 1alpha,25(OH)(2)D(3). In muscle cells, anti-VDR antisense inhibited steroid-induced Ca(2+) and Mn(2+) influx and co-immunoprecipitation of TRPC3 and VDR was observed, suggesting an association between both proteins and a functional role of the receptor in 1alpha,25(OH)(2)D(3) activation of CCE. In osteoblasts, two PCR fragments showing high homology with human INAD-like sequences were obtained. Northern blot and antisense functional assays suggested the involvement of the INAD-like p...Continue Reading

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Citations

Jun 15, 2007·The Journal of Membrane Biology·Graziello GeneauLaurent Cronier
Nov 7, 2006·Microcirculation : the Official Journal of the Microcirculatory Society, Inc·Larissa A ShimodaJ T Sylvester
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Dec 21, 2010·Archives of Biochemistry and Biophysics·Susana MorelliGraciela Santillan
Jun 15, 2011·Molecular and Cellular Endocrinology·Ricardo L Boland
Feb 22, 2012·Current Opinion in Pharmacology·Philippe Gailly

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