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Journal of Neuroscience, Vol 16, 2044-2056, Copyright © 1996 by Society for Neuroscience
Immunocytochemical localization of group III metabotropic glutamate receptors in the hippocampus with subtype-specific antibodies
SR Bradley, AI Levey, SM Hersch and PJ Conn
Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
The metabotropic glutamate receptors (mGluRs) mGluR4 and mGluR7 have been
postulated to serve as presynaptic autoreceptors in the hippocampal
formation. The cellular and synaptic localization of these proteins in the
hippocampus, however, is not known. Polyclonal antibodies that specifically
react with mGluR4a or mGluR7 were produced and used for immunocytochemical
localization of these receptor proteins. Immunoblot analysis with mGluRs
expressed in stably transfected baby hamster kidney cells suggests that
each of the antibodies reacts specifically with the appropriate mGluR
subtype but not with other mGluRs. Both antibodies recognized native
proteins in rat brain with molecular weights similar to the molecular
weights of the bands in mGluR-transfected cell lines. The distribution of
mGluR4a immunoreactivity was fairly uniform in all brain regions.
Immunoreactivity for mGluR7 was variable in different brain regions and
closely paralleled the previously reported distribution of mGluR7 mRNA.
Immunocytochemistry with light and electron microscopy (EM) revealed that
immunoreactivities for mGluR4a and mGluR7 are widely but differentially
distributed throughout the hippocampal formation. Staining with antibodies
directed against mGluR4a was intense in cell bodies and dendrites of
pyramidal cells, granule cells, and scattered interneurons. Analysis at the
EM level revealed postsynaptic mGluR4a localization at asymmetrical
(presumably glutamatergic) synapses and presynaptic localization at both
asymmetrical and symmetrical synapses. Immunoreactivity for mGluR7 revealed
largely presynaptic localization of this receptor at asymmetrical but not
symmetrical synapses. These data are consistent with a largely presynaptic
role of mGluR7 in the hippocampus and suggest that mGluR4 may have both
presynaptic and postsynaptic functions.
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