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PTH-CALCITONIN-VITAMIN D-BONE |
Endocrine Research Unit, Department of Veterans Affairs Medical Center, Department of Medicine, University of California, San Francisco, California 94121
Address all correspondence and requests for reprints to: Dolores Shoback, Endocrine Research Unit, 111N, San Francisco VA Medical Center, 4150 Clement Street, San Francisco, California 94121. E-mail: . dolores{at}itsa.ucsf.edu
Previous studies in chondrogenic RCJ3.1C5.18 (C5.18) cells showed that growth of these cells at high extracellular Ca2+ concentrations ([Ca2+]o) reduced the expression of markers of early chondrocyte differentiation. These studies addressed whether raising [Ca2+]o accelerates C5.18 cell differentiation and whether Ca2+ receptors (CaRs) are involved in coupling changes in [Ca2+]o to cellular responses. We found that high [Ca2+]o increased expression of osteopontin (OP), osteonectin, and osteocalcin, all markers of terminal differentiation, in C5.18 cells and increased the production of matrix mineral. Overexpression of wild-type CaR cDNA in C5.18 cells suppressed proteoglycan synthesis and aggrecan RNA, two early differentiation markers, and increased OP expression. The sensitivity of these parameters to changes in [Ca2+]o was significantly increased, as indicated by left-shifted dose-responses. In contrast, stable expression of a signaling-defective CaR mutant (Phe707Trp CaR) in C5.18 cells, presumably through dominant-negative inhibition of endogenous CaRs, blocked the suppression of aggrecan RNA levels and proteoglycan accumulation and the enhancement of OP expression by high [Ca2+]o. These data support a role for CaRs in mediating high [Ca2+]o-induced differentiation of C5.18 cells.
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