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Volume 16, Number 20, Issue of October 15, 1996 pp. 6353-6363
Copyright ©1996 Society for Neuroscience

Functional Diversity of P-Type and R-Type Calcium Channels in Rat Cerebellar Neurons

Received May 6, 1996; revised July 9, 1996; accepted July 24, 1996.

Angelita Tottene, Alessandra Moretti, and Daniela Pietrobon

Department of Biomedical Sciences and Consiglio Nazionale delle Ricerche Center of Biomembranes, University of Padova, 35131 Padova, Italy

By combining single-channel and whole-cell patch-clamp recordings, we have established the sensitivity to omega -agatoxin IVA and omega -conotoxin MVIIC (SNX-230) of G1, G2, and G3, the three novel non-L-, non-N-type Ca2+ channels characterized previously in rat cerebellar granule cells. G1 channels were blocked irreversibly by both omega -conotoxin MVIIC and low doses of omega -agatoxin IVA (saturation at 50 nM). Thus, according to pharmacological criteria, G1 channels must be classified as P-type Ca2+ channels. Being slowly inactivating during depolarizing pulses and completely inactivated at voltages in which steady-state inactivation of P-type channels in Purkinje cells is negligible, G1 represents a novel P subtype. Neither G2 nor G3 was blocked irreversibly by omega -conotoxin MVIIC, and therefore both are R-type Ca2+ channels. G2 and G3 have some biophysical properties similar to those of low-voltage-activated (LVA) Ca2+ channels (e.g., voltage range for steady-state inactivation, V1/2 = -90 mV), some properties similar to those of high-voltage-activated (HVA) Ca2+ channels (e.g., high sensitivity to Cd2+ block), and other properties intermediate between those of LVA and HVA Ca2+ channels, with LVA properties prevailing in G2 and HVA properties prevailing in G3. The R-type whole-cell current was inhibited by Ni2+ with a biphasic dose-response curve (IC50: 4 and 153 µM), suggesting that G2 and G3 may have a different sensitivity to Ni2+ block. Our results uncover functional diversity of both native P-type and R-type Ca2+ channels and show that R subtypes with distinct biophysical properties are coexpressed in rat cerebellar granule cells.

Key words: calcium channel; cerebellum; granule neuron; patch clamp; conotoxin; agatoxin; channel diversity




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