yo-pro-1 and zinc-chloride

The pH of the airway surface liquid (ASL) plays a pivotal role in maintaining the proper function of the respiratory epithelium. In patients with cystic fibrosis (CF) acidic ASL has been observed. Thus, alkalinization of ASL itself might be beneficial in CF. The aim of this study was to investigate the role of extracellular pH (pH(o)) on the alternative Ca(2+)-activated Cl(-) channels (CaCCs) in CF airway epithelial cells.. The [Ca(2+)](i) and viability of CF airway epithelial cells (IB3-1) were assessed using Fluo-3/AM and YO-PRO-1 fluorescent dyes, respectively. Ion currents were detected in whole-cell configuration using the patch clamp technique.. Extracellular alkalinization (pH(o) 8.2) stimulated Ca(2+) entry and inward currents in low Na(+) containing medium. The inward currents were blocked by the removal of extracellular Ca(2+), chelating cytosolic Ca(2+), as well as by the application of niflumic acid and DIDS. While Zn(2+) promoted sustained Ca(2+) entry in pH(o)-dependent manner, it inhibited the anion conductance. The low external Na(+) concentrations and alkaline pH(o) were well tolerated by the cells.. Stimulation of CaCCs could be achieved by alkalinization of the extracellular environment in CF airway epithelial cells. Zn(2+) directly blocked, however indirectly enhanced the activity of Cl(-) conductance.

Topics: 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; Aniline Compounds; Benzoxazoles; Calcium; Cell Line; Chloride Channels; Chlorides; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Epithelial Cells; Fluorescent Dyes; Humans; Hydrogen-Ion Concentration; Niflumic Acid; Patch-Clamp Techniques; Quinolinium Compounds; Respiratory Mucosa; Xanthenes; Zinc Compounds