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MUSCLE CELL BIOLOGY AND CELL MOTILITY

Activation of Ca²⁺-activated Cl^- current by depolarizing steps in rabbit urethral interstitial cells

Smooth Muscle Group, Department of Physiology, The Queen's University of Belfast, Belfast BT9 7BL, Northern Ireland, United Kingdom

Submitted 6 September 2002 ; accepted in final form 25 March 2003

Interstitial cells were isolated from strips of rabbit urethra for study using the amphotericin B perforated-patch technique. Depolarizing steps to -30 mV or greater activated a Ca²⁺ current (I_Ca), followed by a Ca²⁺-activated Cl^- current, and, on stepping back to -80 mV, large Cl^- tail currents were observed. Both currents were abolished when the cells were superfused with Ca²⁺-free bath solution, suggesting that Ca²⁺ influx was necessary for activation of the Cl^- current. The Cl^- current was also abolished when Ba²⁺ was substituted for Ca²⁺ in the bath or the cell was dialyzed with EGTA (2 mM). The Cl^- current was also reduced by cyclopiazonic acid, ryanodine, 2-aminoethoxydiphenyl borate (2-APB), and xestospongin C, suggesting that Ca²⁺-induced Ca²⁺ release (CICR) involving both ryanodine and inositol 1,4,5-trisphosphate receptors contributes to its activation.

interstitial cells; urethra; calcium-activated chloride current; calcium-induced calcium release; inositol 1,4,5-trisphosphate; ryanodine

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