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Journal of Neuroscience, Vol 7, 1866-1886, Copyright © 1987 by Society for Neuroscience
Resolving GABAA/benzodiazepine receptors: cellular and subcellular localization in the CNS with monoclonal antibodies
JG Richards, P Schoch, P Haring, B Takacs and H Mohler
Monoclonal antibodies, raised against a purified GABAA/benzodiazepine
receptor complex from bovine cerebral cortex, have been used to visualize
the cellular and subcellular distribution of receptorlike immunoreactivity
in the rat CNS, cat spinal cord, and bovine and postmortem human brain. Two
different antibodies have been used for these studies; bd-17 recognizes the
beta-subunit (Mr 55 kDa) in all the species tested, whereas bd-24
recognizes the alpha-subunit (Mr 50 kDa) of bovine and human but not rat
and cat tissues. In bovine and human brain, both antibodies produced very
similar staining patterns, indicating a homogeneous receptor composition,
at least in the brain areas investigated. The general distribution and
density of receptor antigenic sites in all tissues studied were very
similar to that of benzodiazepine binding sites radiolabeled with 3H-Ro
15-1788 and of glutamate decarboxylase (GAD)-stained nerve terminals. The
results demonstrate a very high receptor density (around neuronal cell
bodies and processes or less discretely distributed) in the rat olfactory
bulbs, cerebral cortex, ventral pallidum, islands of Calleja, globus
pallidus, hippocampus, dentate gyrus, substantia nigra, geniculate nuclei,
inferior colliculus, cerebellum, reticular formation, spinal cord, and
retina. In contrast, no receptors could be detected in white matter,
pineal, pituitary, adrenals, and superior cervical ganglia. Only among the
cerebellar layers did we observe a conspicuous difference between the
staining intensity and the radiolabeling. In bovine and postmortem human
brain, e.g., hippocampus, dentate gyrus, cerebral cortex, and substantia
nigra, the same close correlation between the immunohistochemical and
radiohistochemical findings was observed. At the electron microscopic
level, the immune reaction product in the rat substantia nigra and globus
pallidus, for example, was localized to pre- and postsynaptic membranes of
axodendritic and axosomatic synapses. Whether the presynaptic labeling
represents GABA autoreceptors is discussed. In the near future, the
monoclonal antibodies will be used in double-labeling experiments with GAD
to identify those GABAergic projections that are modulated by
benzodiazepine minor tranquillizers. Furthermore, they could also be used,
in studies of postmortem human brain, to diagnose receptor dysfunction
possibly associated with CNS disorders such as epilepsy.
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