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Sensing of Extracellular Cations in CasR-deficient Osteoblasts

J. Biol. Chem., 4 February 2000
Vol. 275, Issue 5, p. 3256-3263
DOI: 10.1074/jbc.275.5.3256

Sensing of Extracellular Cations in CasR-deficient Osteoblasts

EVIDENCE FOR A NOVEL CATION-SENSING MECHANISM*

  1. Min Pi,
  2. Sanford C. Garner,
  3. Patrick Flannery,
  4. Robert F. Spurney and
  5. L. Darryl Quarles
  1. From the Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710

    Abstract

    We isolated osteoblastic cell lines from wild-type (CasR +/+) and receptor null (CasR −/−) mice to investigate whetherCasR is present in osteoblasts and accounts for their responses to extracellular cations. Osteoblasts from bothCasR +/+ and CasR −/−mice displayed an initial period of cell replication followed by a culture duration-dependent increase in alkaline phosphatase activity, expression of osteocalcin, and mineralization of extracellular matrix. In addition, a panel of extracellular cations, including aluminum and the CasR agonists gadolinium and calcium, stimulated DNA synthesis, activated a transfected serum response element-luciferase reporter construct, and inhibited agonist-induced cAMP in CasR −/− osteoblasts. The functional responses to these cations were identical inCasR +/+ and CasR −/−osteoblasts. Thus, the absence of CasR alters neither the maturational profile of isolated osteoblast cultures nor their in vitro responses to extracellular cations. In addition,CasR transcripts could not be detected by reverse transcription-polymerase chain reaction with mouse specific primers in either CasR +/+ orCasR −/− osteoblasts, and immunoblot analysis with a CasR-specific antibody was negative forCasR protein expression in osteoblasts. The presence of a cation-sensing response in osteoblasts fromCasR −/− mice indicates the existence of a novel osteoblastic extracellular cation-sensing mechanism.

    Footnotes

    • * This work was supported in part by Grants R01-AR37308 and R01-AR43468 from NIAMS, National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

      The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) .

    • To whom correspondence should be addressed: Duke University Medical Center, P. O. Box 3036, Durham, NC 27710. Tel.: 919-660-6855; Fax: 919-684-4476; E-mail: Quarl001@mc.duke.edu.

    • Abbreviations:
      RT-PCR

      reverse transcription-polymerase chain reaction

      bp

      base pair(s)

      SRE

      serum response element

      TBS

      Tris-buffered saline

      PGE1

      prostaglandin E1

      • Received July 2, 1999.
      • Revision received September 14, 1999.

    Citation:

    M. Pi, S. C. Garner, P. Flannery, R. F. Spurney, and L. D. Quarles, Sensing of Extracellular Cations in CasR-deficient Osteoblasts. J. Biol. Chem. 275, 3256-3263 (2000).

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