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The Journal of Neuroscience, October 15, 2000, 20(20):7614-7621
G-Protein Types Involved in Calcium Channel Inhibition at a
Presynaptic Nerve Terminal
Rukmini R.
Mirotznik,
Xu
Zheng, and
Elis F.
Stanley
Synaptic Mechanisms Section, National Institute of Neurological
Disorders and Stroke, National Institutes of Health, Bethesda, Maryland
20892
The inhibition of presynaptic calcium channels via
G-protein-dependent second messenger pathways is a key mechanism of
transmitter release modulation. We used the calyx-type nerve terminal
of the chick ciliary ganglion to examine which G-proteins are involved in the voltage-sensitive inhibition of presynaptic N-type calcium channels. Adenosine caused a prominent inhibition of the calcium current that was totally blocked by pretreatment with pertussis toxin
(PTX), consistent with an exclusive involvement of
Go/Gi in the G-protein pathway.
Immunocytochemistry was used to localize these G-protein types to the
nerve terminal and its transmitter release face. We used two approaches
to test for modulation by other G-protein types. First, we treated the
terminals with ligands for a variety of G-protein-linked
neurotransmitter receptor types that have been associated with
different G-protein families. Although small inhibitory effects were
observed, these could all be eliminated by PTX, indicating that in this
terminal the Gi family is the sole transmitter-induced
G-protein inhibitory pathway. Second, we examined the kinetics of
calcium channel inhibition by uncaging the nonselective and
irreversible G-protein activator GTP S, bypassing the receptors. A
large fraction of the rapid GTPS-induced inhibition persisted,
consistent with a Go/Gi-independent
pathway. Immunocytochemistry identified Gq,
G11, G12, and G13 as
potential PTX-insensitive second messengers at this terminal. Thus, our
results suggest that whereas neurotransmitter-mediated calcium channel
inhibition is mainly, and possibly exclusively, via
Go/Gi, other rapid
PTX-insensitive G-protein pathways exist that may involve novel, and
perhaps transmitter-independent, activating mechanisms.
Key words:
nerve terminal; G-protein; G-protein type; calcium
channel; presynaptic; calcium channel modulation; calcium channel
inhibition; transmitter release; synaptic strength; chick; calyx
synapse; chick ciliary ganglion
Copyright © 2000 Society for Neuroscience 0270-6474/00/20207614-08$05.00/0
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