CALCIUM-DEPENDENT ENHANCEMENT OF DEPLETION-ACTIVATED CALCIUM CURRENT IN JURKAT T-LYMPHOCYTES

We have obtained evidence that the Ca2+ selective current activated by Ca2+ store depletion (Ca2+ release-activated Ca2+ current; I-crac) in Jurkat T lymphocytes is augmented in a time-dependent manner by Ca2itself. Whole cell patch clamp experiments employed high cytosolic Ca2 +-buffering conditions to passively deplete Ca2+ stores. Rapidly switc hing,o to nominally Ca2+-free extracellular buffer instantaneously red uced I-crac measured at -100 mV to leak current level. Unexpectedly, r eadmission of 2 mM Ca2+ instantaneously restored only 38 +/- 5% (mean +/- SEM; n = 9) of the full I-crac amplitude. The remainder reappeared in a monotonic time-dependent manner over 10 to 20 sec. Rapid vs. slo w intracellular Ca2+ chelators did not alter this process, and inorgan ic I-crac blockers did not regenerate it, arguing against an intracell ular site of action. The effect was specific to Ca2+: introduction of the permeant ions, Ba2+ or Sr2+, failed to invoke time-dependent I-cra c reappearance. Moreover, equimolar substitution of Ba2+ for Ca2+ init ially produced Ba2+ current of similar magnitude to the full Ca2+ curr ent, but the Ba2+ current decayed monotonically to <50% of its initial amplitude in <20 sec. Conversely, return to Ca2+ produced a time-depe ndent increase in I-crac to its larger Ca2+ permeation level. Thus Ca2 + appears to selectively promote a reversible transition of I-crac tha t results in larger current flux, and at least partially explains the selectivity of this current for Ca2+ over other divalent ions.