Electrophysiological properties of the human N-type Ca2+ channel: I. Channel gating in Ca2+, Ba2+ and Sr2+ containing solutions.

We have characterized the properties of the human N-type Ca2+ channel produced by the stable co-expression of the alpha(1B-1), alpha(2b)delta and beta(1b) subunits. The channel displayed the expected pharmacology with respect to the toxins omega-CTx-GVIA and omega-CTx-MVIIC, which depressed currents in a voltage-independent fashion. We characterized a variety of biophysical properties of the channel under conditions in which either Ca2+, Ba2+ or Sr2+ was the sole extracellular divalent ion. In all three ions, current-voltage relationships revealed that the channel was clearly high-voltage activated. Current activation was significantly slower in Ca2+ than either Sr2+ or Ba2+. Construction of conductance-voltage relationships from tail current measurements indicated that the channel was more high-voltage activated in Ca2+ than in either Sr2+ or Ba2+. The rank order of current amplitude at +4 mV was Ba2+ > Sr2+ > or = Ca2+. Elevation of the extracellular concentration of Ba2+ increased maximal current amplitude and shifted the current-voltage relationship to the right. In all three ions channel inactivation was complex consisting of three distinct exponentials. Recovery from inactivation was slow taking several seconds to reach completion. Steady-state inactivation curves revealed that channel inactivation became detectable at holding potentials of between -101 and -91 mV depending on the permeating species. The rank order of mid-points of steady state inactivation was (most negative) Sr2+ > Ca2+ > Ba2+ (most positive). Deactivation of the N-type Ca2+ channel was voltage-dependent and very fast in all three ions. The deactivation rate in Ba2+ was significantly slower than that in both Ca2+ and Sr2+, however the voltage-dependence of deactivation rate was indistinguishable in all three ions.