Cell calcium levels of normal and cystic fibrosis nasal epithelium

Pediatr Res. 1988 Jul;24(1):79-84. doi: 10.1203/00006450-198807000-00019.

Abstract

To determine whether epithelial ion transport abnormalities in cystic fibrosis (CF) might reflect abnormal regulation of intracellular Ca2+ levels, cytosolic free calcium (Cai2+) was measured using fura-2 or quin2 in suspensions of normal or CF nasal epithelial cells derived from primary cell culture. The basal Cai2+ level measured with fura-2 in CF nasal epithelia was 155 +/- 9 nM (n = 5), a value not significantly different from normal nasal epithelia (143 +/- 16 nM, n = 5). Total cell calcium was measured by atomic absorption spectroscopy and no differences were observed between CF (6.3 +/- 0.5 nmol/mg protein; n = 3) and normal (6.2 +/- 1.2 nmol/mg protein; n = 3) nasal epithelial cells. Placing Na+ loaded cells in a low (10 mM) extracellular Na+ solution resulted in a rapid increase in Cai2+ consistent with Ca2+ uptake via a plasmalemmal Na+-Ca2+ exchanger. The level of Cai2+ achieved by this low Na+ maneuver was not significantly different in CF cells compared to normal cells. Neither isoproterenol (10(-5) M) nor forskolin (10(-6) M) had any effect on Cai2+ in normal or CF nasal epithelial cells. Thus, it appears that differences in cell Cai2+, as measured by fluorescent chelators in suspensions of cultured cells, do not account for the abnormalities in basal or isoproterenol stimulated ion transport in CF tissues.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aminoquinolines
  • Benzofurans
  • Calcium / metabolism*
  • Cells, Cultured
  • Child
  • Cyclic AMP / metabolism
  • Cystic Fibrosis / metabolism*
  • Cytosol / metabolism
  • Epithelium / drug effects
  • Epithelium / metabolism
  • Ethers / pharmacology
  • Female
  • Fluorescent Dyes
  • Fura-2
  • Humans
  • Ionomycin
  • Isoproterenol / pharmacology
  • Male
  • Middle Aged
  • Nasal Mucosa / drug effects
  • Nasal Mucosa / metabolism*
  • Ouabain / pharmacology
  • Sodium / metabolism
  • Spectrophotometry, Atomic

Substances

  • Aminoquinolines
  • Benzofurans
  • Ethers
  • Fluorescent Dyes
  • Ionomycin
  • Ouabain
  • Sodium
  • Cyclic AMP
  • Isoproterenol
  • Quin2
  • Calcium
  • Fura-2