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Cellularand synapticlocalizationof NMDA and non-NMDA
receptorsubunitsin neocortex:organizational features
related to corticalcircuitry, function and disease
George W. Huntley, James C. Vickers and John H. Morrison
George W. Huntley
andJohnH. Mort~son
areat the Fishberg
Research Center for
Neurobiology, Box
1065, The Mount
Sinai Schoolof
Medicine, One
Gustave L. Levy
Place,NY 10029,
USA, andJames C
ricketsis at the Dept
of Pathology, Clinical
School, University of
Tasmania,Hobart,
Tasmania.
Excitatory amino acid (EAA) receptors are an import-
ant component of neocortical circuitry as a result of
their role as the principal mediators of excitatory syn-
@tic activity, as well as their involvement in use-
dependent modifications of synaptic efficacy, excito-
toxicity and cell death. The diversity in the effects
generated by EAA-receptor activation can be attributed
to multiple receptor subOrpes, each of which is composed
of multimeric assemblies of functionally distinct recep-
tor subunits. The use of subunit-specific antibodies and
molecular probes now makes it feasible to localize
individual receptor subunits anatomically with a high
level of cellular and synaptic resolution. Initial studies
of the distribution of immunocytochemically localized
EAA-receptor subunits suggest that particular subunit
combinations exhibit a differential cellular, laminar
and regional distribution in the neocortex. While such
patterns might indicate that the functional heterogeneffy
of EAA-receptor-linked circuits, and the cell types in
which they operate, are based partly on differential
subunit parcellation, a definitive integration of these
anatomical details into current schemes of cortical
circuitry and organization awaits many further studies.
Ideally, such studies should link a high level of mol-
ecular precision regarding subunit localization with
synaptic details of identified connections and neuro-
chemical features of neocortical cells.
Glutamate and aspartate are the excitatory amino acid
(EAA) neurotransmitters of the majority of intrinsic
cerebral cortical neurons (pyramidal cells and spiny
stellate cells) and of thalamocortical relay cells ~. The
excitatory synaptic activity conveyed by these cells is
mediated by three pharmacologically defined subtypes
of ionotropic glutamate receptors: D,L-o~-amino-3-
hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/
low-affinity k.ainate (AMPA receptors), high-affinity
kainate (kalnate receptors) and N-methyl-D-aspartate
(NMDA) receptors. Molecular cloning and functional
expression techniques have demonstrated that each
of the pharmacologically defined EAA-receptor sub-
types is composed of multimeric assemblies of subunit
proteins, details of which have been summarized in
several recent reviews (for example, see Refs 2-4).
Based on common electrophysiological and pharma-
cological properties and similar gene sequences,
distinct subunit families are recognized: subunits
GIuR1-4 compose the AMPA receptors; subunits
GIuR5-7 and the kainate-binding proteins KA1 and
KA2 compose the kainate receptors; and subunits
NMDAR1 and NMDAR2A-D compose the NMDA
receptors. A further level of molecular and functional
diversity arises from the existence of many subunits
in several alternatively spliced forms. In addition,
many subunit transcripts are subject to RNA editing, a
mechanism that appears to be particularly important
for determining subunit properties such as per-
meability to Ca2+, and rectification characteristics 3.
Although the precise subunit composition of EAA
receptors at any specific postsynaptic site is largely
unknown, different combinations of subunits yield
functionally distinct receptor-channels when ex-
pressed in Xenopus oocytes or mammalian cells.
For example, receptors composed of only NMDAR1
subunits yield functional channels displaying low-
amplitude elicited currents, whereas receptors con-
sisting of any of the NMDAR2 subunits without
NMDAR1 do not yield functional channels. However,
combining NMDAR1 with any of the NMDAR2 sub-
units greatly potentiates elicited currents, suggesting
that NMDAR1 might be an obligatory component of all
NMDA receptors4. Moreover, different combinations
of NMDA2A-D with NMDAR1 yield channels that
vary in sensitivity to Mg 2+ block, glycine potentiation,
and pharmacological properties 4. In addition, channels
assembled from combinations of GluR1, GluR3 or
GIuR4 display strong inward rectification and are
permeable to Ca2+ when activated, while the incor-
poration of GluR2 into any combination of AMPA-
receptor subunits yields channels displaying linear or
outwardly rectifying current-voltage (l-V) relation-
ships, and also prevents channel permeability to Ca2+
(Ref. 3).
The anatomical localization of EAA-receptor sub-
types has been principally by autoradiographic
methods5, which are now limited in that individual
~ubnnits or splice variants cannot be distinguished,
nor is resolution at the cellular or synaptic level
possible. In situ hybridization studies in rats
using subunit-specific probes were the first studies to
suggest that the emerging molecular complexity of
EAA subunits and their splice variants is paralleled by
a high degree of anatomical specificity in the distri-
bution of subunits across functionally ,distinct cell
types and regions6-8. The generation of subunit-
specific antibodies and molecular probes makes it
536 © 1994, ElsevierScience Ltd TINS, Vol. 17, NO. 12, 1994