Sensing of extracellular cations in CasR-deficient osteoblasts. Evidence for a novel cation-sensing mechanism

Min Pi, Sanford C. Garner, Patrick Flannery, Robert F. Spurney, Leigh Quarles

Research output: Contribution to journalArticle

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Abstract

We isolated osteoblastic cell lines from wild-type (CasR(+/+)) and receptor null (CasR(-/-)) mice to investigate whether CasR is present in osteoblasts and accounts for their responses to extracellular cations. Osteoblasts from both CasR(+/+) 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 in CasR(+/+) 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(+/+) or CasR(-/-) osteoblasts, and immunoblot analysis with a CasR-specific antibody was negative for CasR protein expression in osteoblasts. The presence of a cation-sensing response in osteoblasts from CasR(-/-) mice indicates the existence of a novel osteoblastic extracellular cation-sensing mechanism.

Original languageEnglish (US)
Pages (from-to)3256-3263
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number5
DOIs
StatePublished - Feb 4 2000
Externally publishedYes

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Osteoblasts
Cations
Serum Response Element
Polymerase chain reaction
Osteocalcin
Gadolinium
Transcription
Luciferases
Aluminum
Cell culture
Reverse Transcription
Extracellular Matrix
Alkaline Phosphatase
Cells
Calcium
Cell Line
Polymerase Chain Reaction
Antibodies
DNA

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sensing of extracellular cations in CasR-deficient osteoblasts. Evidence for a novel cation-sensing mechanism. / Pi, Min; Garner, Sanford C.; Flannery, Patrick; Spurney, Robert F.; Quarles, Leigh.

In: Journal of Biological Chemistry, Vol. 275, No. 5, 04.02.2000, p. 3256-3263.

Research output: Contribution to journalArticle

Pi, Min ; Garner, Sanford C. ; Flannery, Patrick ; Spurney, Robert F. ; Quarles, Leigh. / Sensing of extracellular cations in CasR-deficient osteoblasts. Evidence for a novel cation-sensing mechanism. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 5. pp. 3256-3263.
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