Calcimimetic R-568 effects on activity of R990G polymorphism of calcium-sensing receptor

Annalisa Terranegra, Anita Ferraretto, Elena Dogliotti, Milena Scarpellini, Sabrina Corbetta, Anna Maria Barbieri, Anna Spada, Teresa Arcidiacono, Francesco Rainone, Andrea Aloia, Daniele Cusi, Giuseppe Vezzoli, Laura Soldati

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Previous studies have demonstrated a gain-of-function of the calcium-sensing receptor (CASR) gene R990G polymorphism. In this study, activation of the R990G CASR stably transfected in HEK-293 (HEK-990G) cells compared with that of the common variant (HEK-wild-type (WT)) by increasing concentrations of CaCl2 or calcimimetic R-568 caused significantly higher intracellular free calcium concentration ([Ca2+]i) and lower Ca-EC50. Moreover, the [Ca2+]i oscillation percentage was higher with a larger sinusoidal pattern in HEK-990G. R-568 induced a shift of the oscillatory events from 4 to 2 mmol/l extracellular calcium concentration in HEK-990G cells and increased the sinusoidal oscillation percentage in comparison with HEK-WT. Preincubation with thapsigargin or phospholipase C inhibitors completely prevented oscillations in both cell lines, consistent with the involvement of the inositol trisphosphate pathway, while protein kinase C inhibitor prevented oscillations in HEK-WT cells only. Finally, CaCl2 and R-568 caused a significant increase in p44/42 extracellular signaling-regulated kinase phosphorylation, with the mean Ca-EC50 values being significantly lower in HEK-990G. Our findings demonstrated that the 990G allele is associated with high sensitivity to R-568, which provided new evidence for differences in CASR signaling.

Original languageEnglish
Pages (from-to)245-256
Number of pages12
JournalJournal of Molecular Endocrinology
Volume45
Issue number4
DOIs
Publication statusPublished - Oct 2010
Externally publishedYes

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N-(2-chlorophenylpropyl)-1-(3-methoxyphenyl)ethylamine
Calcium-Sensing Receptors
Calcium
Thapsigargin
Protein C Inhibitor
HEK293 Cells
Type C Phospholipases
Inositol
Protein Kinase Inhibitors
Protein Kinase C
Phosphotransferases
Alleles
Phosphorylation
Cell Line
Genes

ASJC Scopus subject areas

  • Endocrinology
  • Molecular Biology

Cite this

Terranegra, A., Ferraretto, A., Dogliotti, E., Scarpellini, M., Corbetta, S., Barbieri, A. M., ... Soldati, L. (2010). Calcimimetic R-568 effects on activity of R990G polymorphism of calcium-sensing receptor. Journal of Molecular Endocrinology, 45(4), 245-256. https://doi.org/10.1677/JME-10-0034

Calcimimetic R-568 effects on activity of R990G polymorphism of calcium-sensing receptor. / Terranegra, Annalisa; Ferraretto, Anita; Dogliotti, Elena; Scarpellini, Milena; Corbetta, Sabrina; Barbieri, Anna Maria; Spada, Anna; Arcidiacono, Teresa; Rainone, Francesco; Aloia, Andrea; Cusi, Daniele; Vezzoli, Giuseppe; Soldati, Laura.

In: Journal of Molecular Endocrinology, Vol. 45, No. 4, 10.2010, p. 245-256.

Research output: Contribution to journalArticle

Terranegra, A, Ferraretto, A, Dogliotti, E, Scarpellini, M, Corbetta, S, Barbieri, AM, Spada, A, Arcidiacono, T, Rainone, F, Aloia, A, Cusi, D, Vezzoli, G & Soldati, L 2010, 'Calcimimetic R-568 effects on activity of R990G polymorphism of calcium-sensing receptor', Journal of Molecular Endocrinology, vol. 45, no. 4, pp. 245-256. https://doi.org/10.1677/JME-10-0034
Terranegra, Annalisa ; Ferraretto, Anita ; Dogliotti, Elena ; Scarpellini, Milena ; Corbetta, Sabrina ; Barbieri, Anna Maria ; Spada, Anna ; Arcidiacono, Teresa ; Rainone, Francesco ; Aloia, Andrea ; Cusi, Daniele ; Vezzoli, Giuseppe ; Soldati, Laura. / Calcimimetic R-568 effects on activity of R990G polymorphism of calcium-sensing receptor. In: Journal of Molecular Endocrinology. 2010 ; Vol. 45, No. 4. pp. 245-256.
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