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The Journal of Neuroscience, June 15, 1999, 19(12):4755-4761
Sequestration of G-Protein   Subunits by Different G-Protein
 Subunits Blocks Voltage-Dependent Modulation of Ca2+
Channels in Rat Sympathetic Neurons
Seong-Woo
Jeong and
Stephen R.
Ikeda
Laboratory of Molecular Physiology, Guthrie Research Institute,
Sayre, Pennsylvania 18840
The membrane-delimited and voltage-dependent inhibition of N-type
Ca2+ channels is mediated by G  subunits.
Previously, exogenous excess GDP-bound G oA has been
shown to dramatically attenuate the norepinephrine (NE)-mediated
Ca2+ current inhibition by sequestration of G
subunits in rat superior cervical ganglion (SCG) neurons. In the
present study, we determined whether the attenuation of NE-mediated
modulation is specific to GoA or shared by a number of
closely related (Gtr, GoB, Gi1, Gi2,
Gi3, Gz) or unrelated
(Gs, Gq,
G11, G16,
G12, G13) G subunits.
Individual G subunits from different subfamilies were transiently
overexpressed in SCG neurons by intranuclear injection of mammalian
expression vectors encoding the desired protein. Strikingly, all G
subunits except Gz nearly blocked basal facilitation and
NE-mediated modulation. Likewise, VIP-mediated Ca2+
current inhibition, which is mediated by cholera toxin-sensitive G-protein, was also completely suppressed by a number of G subunits overexpressed in neurons. Gs expression produced either
enhancement or attenuation of the VIP-mediated modulation an effect
that seemed to depend on the expression level. The onset of the
nonhydrolyzable GTP analog, guanylylimidodiphosphate-mediated
facilitation was significantly delayed by overexpression of different
GDP-bound G subunits. Taken together, these data suggest that a wide
variety of G subunits are capable of forming heterotrimers with
endogenous G subunits mediating voltage-dependent
Ca2+ channel inhibition. In conclusion, coupling
specificity in signal transduction is unlikely to arise as a result of
restricted G/G interaction.
Key words:
calcium channels; G subunit; G subunit; G-proteins; voltage-dependent inhibition; intranuclear injection; sympathetic neuron; coupling specificity
Copyright © 1999 Society for Neuroscience 0270-6474/99/19124755-07$05.00/0
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