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The Journal of Neuroscience, August 1, 2000, 20(15):5623-5629

A Voltage-Independent Calcium Current Inhibitory Pathway Activated by Muscarinic Agonists in Rat Sympathetic Neurons Requires Both G_q/11 and G

Paul J. Kammermeier, Victor Ruiz-Velasco, and Stephen R. Ikeda

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

Calcium current modulation by the muscarinic cholinergic agonist oxotremorine methiodide (oxo-M) was examined in sympathetic neurons from the superior cervical ganglion of the rat. Oxo-M strongly inhibited calcium currents via voltage-dependent (VD) and voltage-independent (VI) pathways. These pathways could be separated with the use of the specific M₁ acetylcholine receptor antagonist M₁-toxin and with pertussis toxin (PTX) treatment. Expression by nuclear cDNA injection of the regulator of G-protein signaling (RGS2) or a phospholipase C1 C-terminal construct (PLC-ct) selectively reduced VI oxo-M modulation in PTX-treated and untreated cells. Expression of the G buffers transducin (G_tr) and a G-protein-coupled-receptor kinase (GRK3) construct (MAS-GRK3) eliminated oxo-M modulation. Activation of the heterologously expressed neurokinin type 1 receptor, a G_q/11-coupled receptor, resulted in VI calcium current modulation. This modulation was eliminated with coexpression of G_tr or MAS-GRK3. Cells expressing G₁₂ were tonically inhibited via the VD pathway. Application of oxo-M to these cells produced VI modulation and reduced the amount of current inhibited via the VD pathway. Together, these results confirm the requirement for G in VD modulation and implicate G_q-GTP and G as components in the potentially novel VI pathway.

Key words: N-type calcium channel; ion channel modulation; voltage dependent; sympathetic neurons; SCG; G-protein

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Copyright © 2000 Society for Neuroscience  0270-6474/00/20155623-07$05.00/0

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