Interaction between capacitative Ca2+ influx and Ca2+-dependent Cl- currents in Xenopus oocytes.

The relationship between capacitative Ca2+ influx and activation of Ca2+-dependent Cl- channels was monitored in intact Xenopus oocytes following stimulation of 5-hydroxytryptamine (5-HT) receptors, through the activity of Ca2+-dependent Cl- channels using the double-electrode voltage-clamp technique. Under voltage-clamp conditions, 5-HT evoked a rapid transient inward current followed by a slowly developing secondary inward current. The secondary current reflected depletion-activated Ca2+ entry. Hyperpolarising pulses evoked sustained Ca2+-dependent Cl- currents when applied during the transient inward current, but evoked hump-like currents which inactivated rapidly when applied during the secondary inward current. Hump currents arose from Ca2+ entering through the depletion-activated pathway. The hump currents inactivated with hyperpolarising pulses at < 5-s intervals, and recovered monoexponentially with a time constant of around 8 s. Currents in response to hyperpolarising pulses during the transient current did not inactivate, suggesting that inactivation was associated with Ca2+ entry. When ca2+ release evoked by inositol 1,4,5-triphosphate [ins(1,4,5)p3] was prevented by heparin injection, hyperpolarising pulses during ca2+ ionophore application also generated hump currents that were dependent on external ca2+, inactivated and recovered from inactivation with a similar time course as the humps following 5-ht treatment. Pretreatment with the Ca2+ adenosine 5'-triphosphatase (Ca2+ATPase) inhibitor thapsigargin reduced the rate of rise of the hump current, increased the time-to-peak of the current and slowed the rate of decay. Pharmacological interventions to disrupt the cytoskeleton reduced the amplitude of the hump current. It is suggested that, following hyperpolarisation in the presence of Ca2+ entry, the ensuing Ca2+ influx interacts with Cl- channels in a way that might reflect both Ca2+ inhibition of Ca2+ entry and clustering of Cl- channels in the plasma membrane.