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Characterization of cell volume-sensitive chloride currents in freshly prepared and cultured pancreatic acinar cells from early postnatal rats

Andreas Schmid, Robert Blum and Elmar Krause

In freshly prepared and cultured exocrine pancreatic acinar cells from 5- to 7-day-old rats a chloride-selective membrane conductance could be activated by intracellular application of GTPS (40-100 µM), by application of positive pressure (5 cmH₂O) to the pipette interior or by challenging the cells with a hyposmolar bath solution. Hyperosmolar bath solutions inhibited the cell volume-sensitive chloride currents.

The anion permeability sequence of the cell volume-sensitive chloride conductance was I^- > Cl^- Br^- > F^- > methanesulphonate^- > glutamate^-. I^- had a higher permeability but lower conductance than Cl^-. The permeability ratio for P_glutamate/P_Cl was 0·12.

The cell volume-sensitive chloride conductance showed outward rectification. Membrane depolarization to high positive voltages ( +60 mV) caused a time-dependent decay in outward currents.

DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid) and SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulphonic acid) reversibly inhibited the cell volume-sensitive chloride current in a voltage-dependent manner. NPPB (5-nitro-2-(3-phenylpropylamino)-benzoic acid), quinidine, quinine and tamoxifen caused voltage-independent current inhibition.

Combined fura-2 and whole-cell current measurements showed that activation of the cell volume-sensitive chloride current does not involve cytosolic Ca²⁺ signals. Furthermore, there is no evidence that Ca²⁺-activated chloride currents play a significant role in cultured pancreatic acinar cells from 5- to 7-day-old rats.

Polymerase chain reaction followed by DNA sequence analysis indicated the presence of mRNA homologous to the ClC-3 chloride channel in pancreatic tissue from 5-day-old rats.

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