Swelling-induced chloride currents in neuroblastoma cells are calcium dependent

Abstract

The effects of osmotic stress on chloride (Cl-) currents in the human neuroblastoma cell line CHP-100 were evaluated. Following exposure to hypoosmotic solution, an increase in whole-cell Cl- current was observed. This current was blocked by the Cl- channel blocker 5-nitro-2-(3- phenylpropylamino)-benzoic acid (NPPB). In cells loaded with the Cl- permeability marker 125I, exposure to hypoosmotic solution increased 125I efflux by 197 ± 14% (n = 41, p < 0.05) over controls. This increase was sensitive to NPPB. Hypoosmotic stress also increased cytosolic calcium levels (Ca2+) in fura-2-loaded cells. Pretreatment with EGTA inhibited the increase in cytosolic Ca2+, 125I efflux, and whole-cell Cl- current produced by hypoosmotic solution. Antagonists of N-, L-, and T-type Ca2+ channels did not alter stimulation in 125I efflux or cytosolic Ca2+ levels during osmotic stress. However, ω-conotoxin MVIIC, a P-type Ca2+ channel blocker, inhibited hypoosmotically activated whole-cell Cl- currents and increases in cytosolic Ca2+. It is concluded that a Ca2+-dependent change in Cl- permeability is activated in CHP-100 cells in response to osmotic stress. Show more