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The Journal of Neuroscience, January 1, 2000, 20(1):171-178

_1E Subunits Form the Pore of Three Cerebellar R-Type Calcium Channels with Different Pharmacological and Permeation Properties

Angelita Tottene¹, Stephen Volsen², and Daniela Pietrobon¹

¹ Department of Biomedical Sciences, University of Padova, 35121 Padova, Italy, and ² Lilly Research Center, Eli Lilly Company Limited, Windlesham, Surrey GU20 6PH, United Kingdom

R-type Ca²⁺ 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 _1E subunit. However, R-type Ca²⁺ currents with permeation and/or pharmacological properties different from those of recombinant Ca²⁺ channels containing _1E subunits have been described, and therefore the molecular nature of R-type Ca²⁺ channels remains not completely settled. Here, we show that the R-type Ca²⁺ current of rat cerebellar granule cells consists of two components inhibited with different affinity by the _1E selective antagonist SNX482 (IC₅₀ 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 _1E channels; it is larger with Ca²⁺ than with Ba²⁺ as charge carrier, and it is highly sensitive to Ni²⁺ 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 _1A and _1B channels; it is larger with Ba²⁺ than with Ca²⁺ as charge carrier_, and it has a low sensitivity to Ni²⁺ 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 _1E gene, suggesting the existence of _1E isoforms with different pore properties.

Key words: calcium channel; _1E subunit; antisense oligonucleotides; cerebellum; granule cells; permeation; toxin-resistant calcium current

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Copyright © 2000 Society for Neuroscience  0270-6474/0/201171-08$05.00/0

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