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Volume 17, Number 12,
Issue of June 15, 1997
pp. 4672-4687
Copyright ©1997 Society for Neuroscience
Transporters Buffer Synaptically Released Glutamate on a
Submillisecond Time Scale
Received Feb. 4, 1997; revised April 7, 1997; accepted April 8, 1997.
Jeffrey S. Diamond and
Craig E. Jahr
The Vollum Institute, Oregon Health Sciences University,
Portland, Oregon 97201
The role of transporters in clearing free glutamate from the
synaptic cleft was studied in rat CA1 hippocampal neurons cultured on
glial microislands. The time course of free glutamate in the cleft
during a synaptic event was estimated by measuring the extent to which
the rapidly dissociating AMPA receptor antagonist kynurenate (KYN) was
replaced by glutamate during a synaptic response. Dose inhibition of
the AMPA receptor EPSC by KYN was less than predicted by the
equilibrium affinity of the antagonist, and the rise time of AMPA
receptor miniature EPSCs (mEPSCs) was slowed by KYN. Both results
indicated that KYN dissociated from AMPA receptors and was replaced by
synaptically released transmitter. When transporters were blocked by
D,L-threo- -hydroxyaspartic acid
(THA) or Li+, the mEPSC rise time in the presence of
KYN was slowed further, indicating that transporters affect the
glutamate concentration in the first few hundred microseconds of the
synaptic response.
The glutamate transient necessary to cause these effects was determined
by developing a detailed kinetic model of the AMPA receptor. The model
replicated the effects of KYN on the amplitude and rise time of the
synaptic responses when driven by glutamate transients that were
similar to previous estimates (; ).
The effects of THA were replicated by slowing and enlarging the slower
phase of the dual component transient by about 20% or by prolonging
the single component by almost 40%. Because transport is too slow to
account for these effects, it is concluded that transporters buffer
glutamate in the synaptic cleft.
Key words:
glutamate transporters;
AMPA receptor;
kinetic
model;
cultured hippocampal neurons;
buffered diffusion;
miniature
EPSC;
kynurenate;
D,L-threo--hydroxyaspartic acid
(THA)
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F. L. Kidd and J. T. R. Isaac
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S. Eskandari, M. Kreman, M. P. Kavanaugh, E. M. Wright, and G. A. Zampighi
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M. L. Beckman and M. W. Quick
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J. S. Diamond and C. E. Jahr
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M. M. Bolton, A. J. Pittman, and D. C. Lo
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T. S. Otis and M. P. Kavanaugh
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J. E. Hanson and Y. Smith
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Y. Zhang and B. I. Kanner
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R. Rao-Mirotznik, G. Buchsbaum, and P. Sterling
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M. V. Jones, Y. Sahara, J. A. Dzubay, and G. L. Westbrook
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K. P. Lehre and N. C. Danbolt
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J. I. Wadiche and M. P. Kavanaugh
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T. S. Otis and C. E. Jahr
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