Ca(2+)-dependent inactivation of high-threshold Ca(2+) currents in hippocampal granule cells of patients with chronic temporal lobe epilepsy.

ABSTRACT

Intracellular Ca(2+) represents an important trigger for various second-messenger mediated effects. Therefore a stringent control of the intracellular Ca(2+) concentration is necessary to avoid excessive activation of Ca(2+)-dependent processes. Ca(2+)-dependent inactivation of voltage-dependent calcium currents (VCCs) represents an important negative feedback mechanism to limit the influx of Ca(2+) that has been shown to be altered in the kindling model of epilepsy. We therefore investigated the Ca(2+)-dependent inactivation of high-threshold VCCs in dentate granule cells (DGCs) isolated from the hippocampus of patients with drug-refractory temporal lobe epilepsy (TLE) using the patch-clamp method. Ca(2+) currents showed pronounced time-dependent inactivation when no extrinsic Ca(2+) buffer was present in the patch pipette. In addition, in double-pulse experiments, Ca(2+) entry during conditioning prepulses caused a reduction of VCC amplitudes elicited during a subsequent test pulse. Recovery from Ca(2+)-dependent inactivation was slow and only complete after 1 s. Ca(2+)-dependent inactivation could be blocked either by using Ba(2+) as a charge carrier or by including bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) or EGTA in the intracellular solution. The influence of the cytoskeleton on Ca(2+)-dependent inactivation was investigated with agents that stabilize and destabilize microfilaments or microtubules, respectively. From these experiments, we conclude that Ca(2+)-dependent inactivation in human DGCs involves Ca(2+)-dependent destabilization of both microfilaments and microtubules. In addition, the microtubule-dependent pathway is modulated by the intracellular concentration of GTP, with lower concentrations of guanosine triphosphate (GTP) causing increased Ca(2+)-dependent inactivation. Under low-GTP conditions, the amount of Ca(2+)-dependent inactivation was similar to that observed in the kindling model. In summary, Ca(2+)-dependent inactivation was present in patients with TLE and Ammon's horn sclerosis (AHS) and is mediated by the cytoskeleton similar to rat pyramidal neurons. The similarity to the kindling model of epilepsy may suggest the possibility of altered Ca(2+)-dependent inactivation in patients with AHS.