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The Journal of Neuroscience, March 15, 2000, 20(6):2183-2191

Multiple G-Protein Combinations Produce Voltage-Dependent Inhibition of N-Type Calcium Channels in Rat Superior Cervical Ganglion Neurons

Victor Ruiz-Velasco and Stephen R. Ikeda

Laboratory of Molecular Physiology, Guthrie Research Institute, Sayre, Pennsylvania 18840

Activation of several G-protein-coupled receptors leads to voltage-dependent (VD) inhibition of N- and P/Q-type Ca²⁺ channels via G-protein subunits (G). The purpose of the present study was to determine the ability of different G combinations to produce VD inhibition of N-type Ca²⁺ channels in rat superior cervical ganglion neurons. Various G combinations were heterologously overexpressed by intranuclear microinjection of cDNA and tonic VD Ca²⁺ channel inhibition evaluated using the whole-cell voltage-clamp technique. Overexpression of G1-G5, in combination with several different G subunits, resulted in tonic VD Ca²⁺ channel inhibition. Robust Ca²⁺ channel modulation required coexpression of both G and G. Expression of either subunit alone produced minimal effects. To substantiate the apparent lack of G specificity, we examined whether heterologously expressed G displaced native G from heterotrimeric complexes. To this end, mutant G subunits were constructed that differentially modulated N-type Ca²⁺ and G-protein-gated inward rectifier K⁺ channels. Results from these studies indicated that significant displacement does not occur, and thus the observed G modulation can be attributed directly to the heterologously expressed G combinations.

Key words: G-protein; N-type Ca²⁺ channel; GIRK channel; G; ion channel modulation; SCG neurons

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Copyright © 2000 Society for Neuroscience  0270-6474/00/2062183-09$05.00/0

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