Sodium cyanide increases cytosolic free calcium: Evidence for activation of the reversed mode of the Na+/Ca2+ exchanger and Ca2+ mobilization from inositol trisphosphate-insensitive pools

Juliann G. Kiang, Robert C. Smallridge

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

This study characterized the cytosolic free Ca2+ concentration ([Ca2+]i) in NaCN-treated human A-431 cells. The resting [Ca2+]i was 85 ± 8 nM (n = 141) in untreated cells at 37°C, determined with the fura-2 fluorescence probe, When cells were treated with NaCN, [Ca2+]i increased in a time- and NaCN concentration-dependent manner. When cells were exposed to 10 mM NaCN for 10 min, [Ca2+]i increased 278 ± 28% (n = 5) but returned to normal within 45 min after treatment. The [Ca2+]i increase depended on the presence of external Ca2+. La3+ and Cd2+, but not verapamil or nifedipine, inhibited the NaCN-induced [Ca2+]i increase. The NaCN-induced [Ca2+]i increase also depended on external Na+ (K1 2 = 85 mM). The intracellular Na+ concentration, measured with the fluorescence probe SBFI, increased 267 ± 16% after NaCN treatment. The NaCN-induced [Ca2+]i increase was modulated by treatment with ouabain or veratridine and was completely blocked by tetrodotoxin, amiloride (K1 2 = 5.4 μM), and dichlorobenzamil (K1 2 = 0.28 μM). These results suggest NaCN activates the Na+/Ca2+ exchange system. TMB-8 and ryanodine both partially blocked the increase in [Ca2+]i in the presence of external Ca2+, indicating that Ca2+ release from intracellular pools also occurred after the initial Ca2+ influx. NaCN decreased inositol trisphosphates production. U-73122, bradykinin, or monensin did not prevent the NaCN-induced increase in [Ca2+]i. However, the magnitude of the [Ca2+]i increase caused by NaCN was abolished in ionomycin-treated cells, indicating that intracellular Ca2+ release induced by NaCN is derived from an ionomycin-sensitive Ca2+ pool. The results suggest that NaCN initially increased Na+ influx, which activated the reverse mode of a Na+/Ca2+ exchanger, leading to an increase in Ca2+ influx. The Ca2+ influx induced a Ca2+ mobilization from only an ionomycin-sensitive intracellular Ca2+ pool containing ryanodine receptors.

Original languageEnglish (US)
Pages (from-to)173-181
Number of pages9
JournalToxicology and Applied Pharmacology
Volume127
Issue number2
DOIs
StatePublished - Aug 1994

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

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