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Journal of Neuroscience, Vol 13, 2965-2981, Copyright © 1993 by Society for Neuroscience
Selective distribution of kainate receptor subunit immunoreactivity in monkey neocortex revealed by a monoclonal antibody that recognizes glutamate receptor subunits GluR5/6/7
GW Huntley, SW Rogers, T Moran, W Janssen, N Archin, JC Vickers, K Cauley, SF Heinemann and JH Morrison
Fishberg Research Center for Neurobiology, Mount Sinai School of Medicine, New York, New York 10029.
A monoclonal antibody (4F5) was generated against a portion of the putative
extracellular domain of glutamate receptor subunit GluR5. Western blot
analyses and immunocytochemistry of transfected human embryonic kidney 293
cells confirmed that monoclonal antibody 4F5 was specific for GluR5, -6,
and -7 (the three identified members of the kainate receptor subunit
class), but did not recognize GluR1, -2, or -3 (the AMPA/kainate receptor
subunit class). The antibody was subsequently used to examine
immunocytochemically the regional, laminar, and cellular distribution of
GluR5/6/7 receptor subunits at the light and electron microscopic levels in
monkey neocortex. Receptor subunit immunoreactivity was present throughout
all cortical areas examined, but exhibited marked cellular, laminar, and
regional specificity. Typically, pyramidal cell somata and apical dendrites
were well stained. Electron microscopy revealed an extensive cytoplasmic
localization of GluR5/6/7 immunoprecipitate, with intense staining of many
postsynaptic densities, all of which were associated with asymmetric
synapses located on dendritic shafts or dendritic spines. There was no
evidence of stained glial cells or presynaptic axon terminals. In most
areas, labeled cells and dendrites were concentrated in layers II, III, and
V while layers I, IV, and VI typically possessed the fewest and/or least
intensely stained elements. A consistent feature in many areas was groups
of clustered layer V pyramidal cells and bundles of ascending apical
dendrites. Regionally, motor areas and higher-order association areas of
the frontal, parietal, and occipital lobes were more densely stained than
primary sensory areas (somatic sensory and visual cortex), which was
confirmed quantitatively. These data indicate a high degree of selectivity
in the distribution of kainate receptors composed of GluR5/6/7 subunits,
and suggest that functional specificity and diversity in the ubiquitous
excitatory amino acid-utilizing axonal systems in neocortex are achieved in
part by the differential association of particular glutamate receptor
subunits with specific cortical circuits. In addition, the regional,
laminar, and morphological characteristics of GluR5/6/7-immunoreactive
neurons bear a strong similarity to those of the neocortical neurons with
heightened vulnerability in certain neurodegenerative disorders.
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