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Journal of Neuroscience, Vol 13, 3041-3050, Copyright © 1993 by Society for Neuroscience
Modulation of calcium currents by a metabotropic glutamate receptor involves fast and slow kinetic components in cultured hippocampal neurons
Y Sahara and GL Westbrook
Vollum Institute, Oregon Health Sciences University, Portland 97201.
The modulation of high-threshold Ca2+ currents by the selective
metabotropic glutamate receptor (mGluR) agonist (1S,3R)-1-
aminocyclopentane-1,3-dicarboxylic acid (ACPD), was investigated in
cultured hippocampal neurons using whole-cell voltage-clamp recording. ACPD
reduced high-threshold Ca2+ currents carried by Ba2+ with an EC50 of 15.5
microM. The inhibition was reversible, voltage dependent, and blocked by
L-2-amino-3-phosphonopropionic acid (1 mM) or by pretreatment with
pertussis toxin. Inhibition by ACPD was greatly enhanced, and became
irreversible, when the nonhydrolyzable GTP analog GTP gamma S was included
in the whole-cell pipette. In some neurons, the Ba2+ current was inhibited
by L(+)-2-amino-4-phosphonobutanoic acid (L-AP4) as well as ACPD while most
cells were insensitive to L-AP4, suggesting that these agonists activate
distinct receptors. The inhibition of Ca2+ currents was reduced but not
eliminated in the presence of either omega-conotoxin GVIA or nifedipine,
suggesting that both N- and L-type Ca2+ currents were affected. The degree
and kinetics of inhibition were dependent on intracellular calcium. With
[Ca]i < 1 nM, inhibition had a fast onset (t approximately 1-2 sec) and
a rapid recovery, consistent with a membrane-delimited pathway. However, a
slow component of inhibition appeared when the steady state [Ca]i was
increased to 100 nM (t onset approximately 3 min). The slow component did
not require transient Ca2+ influx or release of intracellular Ca2+. We
suggest that Ca2+ channel modulation by ACPD involves either two mGluR
subtypes with separate coupling mechanisms or a single mGluR that couples
to both mechanisms.
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