Cytosolic free Ca2+ concentration exhibits a characteristic temporal pattern during in vitro cartilage differentiation

A possible regulatory role of calcineurin in Ca-signalling of chondrogenic cells

Csaba Matta, János Fodor, Zsolt Szíjgyártó, Tamás Juhász, P. Gergely, L. Csernoch, Róza Zákány

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

We measured changes of cytosolic Ca2+ concentration during chondrogenesis, which occurs in high-density cultures (HDC) of chondrifying chicken mesenchymal cells. A significant, transient elevation was detected in Fura-2-loaded cells on day 3 of culturing, when majority of chondrogenic cells of HDC become differentiated. This 140 nM peak of cytosolic Ca2+ concentration is a result of increased Ca-influx and is indispensable to proper chondrogenesis, because addition of 0.8 mM EGTA to culture medium on day 2 or 3 significantly decreased the intracellular Ca2+ concentration abolishing the Ca2+-peak of day 3 and inhibited cartilage formation. Uncontrolled Ca2+ influx evoked by a Ca2+ ionophore exerted dual effects on chondrogenesis in a concentration-dependent manner; 0.1 mg/L A23187 increased, whereas 5 mg/L A23187 almost totally blocked cartilage formation. Intracellular Ca-stores seemed not to have any significant participation in the regulation of changes of cytosolic Ca2+ concentration of chondrifying cells. Activity of Ca-calmodulin-dependent protein phosphatase, calcineurin responded to changes of intracellular Ca2+ concentration induced by EGTA or A23187 in a differentiation stage-dependent manner. Since inhibition of calcineurin with cyclosporine A eliminated the peak in the cytosolic Ca2+ concentration, an active regulatory role of calcineurin on Ca2+ influx of chondrifying cells can be supposed.

Original languageEnglish
Pages (from-to)310-323
Number of pages14
JournalCell Calcium
Volume44
Issue number3
DOIs
Publication statusPublished - Sep 2008

Fingerprint

Calcineurin
Chondrogenesis
Cartilage
Egtazic Acid
Fura-2
Phosphoprotein Phosphatases
Ionophores
Calcimycin
Calmodulin
Cyclosporine
Culture Media
Chickens
Cell Count
In Vitro Techniques

Keywords

  • Chondrogenesis
  • Cyclosporine A
  • Fura-2
  • High-density culture
  • Intracellular Ca concentration
  • Sox9

ASJC Scopus subject areas

  • Cell Biology
  • Endocrinology
  • Molecular Biology
  • Physiology

Cite this

Cytosolic free Ca2+ concentration exhibits a characteristic temporal pattern during in vitro cartilage differentiation : A possible regulatory role of calcineurin in Ca-signalling of chondrogenic cells. / Matta, Csaba; Fodor, János; Szíjgyártó, Zsolt; Juhász, Tamás; Gergely, P.; Csernoch, L.; Zákány, Róza.

In: Cell Calcium, Vol. 44, No. 3, 09.2008, p. 310-323.

Research output: Contribution to journalArticle

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