α1ESubunits Form the Pore of Three Cerebellar R-Type Calcium Channels with Different Pharmacological and Permeation Properties

R-type Ca(2+) channels cooperate with P/Q- and N-type channels to control neurotransmitter release at central synapses. The leading candidate as pore-forming subunit of R-type channels is the alpha(1E) subunit. However, R-type Ca(2+) currents with permeation and/or pharmacological properties different from those of recombinant Ca(2+) channels containing alpha(1E) subunits have been described, and therefore the molecular nature of R-type Ca(2+) channels remains not completely settled. Here, we show that the R-type Ca(2+) current of rat cerebellar granule cells consists of two components inhibited with different affinity by the alpha(1E) selective antagonist SNX482 (IC(50) values of 6 and 81 nM) and a third component resistant to SNX482. The SNX482-sensitive R-type current shows the unique permeation properties of recombinant alpha(1E) channels; it is larger with Ca(2+) than with Ba(2+) as charge carrier, and it is highly sensitive to Ni(2+) block and has a voltage-dependence of activation consistent with that of G2 channels with unitary conductance of 15 pS. On the other hand, the SNX482-resistant R-type current shows permeation properties similar to those of recombinant alpha(1A) and alpha(1B) channels; it is larger with Ba(2+) than with Ca(2+) as charge carrier(,) and it has a low sensitivity to Ni(2+) block and a voltage-dependence of activation consistent with that of G3 channels with unitary conductance of 20 pS. Gene-specific knock-down by antisense oligonucleotides demonstrates that the different cerebellar R-type channels are all encoded by the alpha(1E) gene, suggesting the existence of alpha(1E) isoforms with different pore properties.