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Journal of Neuroscience, Vol 7, 88-100, Copyright © 1987 by Society for Neuroscience
Corticotropin-releasing factor receptors in the rat central nervous system: characterization and regional distribution
EB De Souza
A stable, iodine-125-labeled analog of rat/human corticotropin- releasing
factor (CRF) was used to define the characteristics of CRF receptors in a
crude mitochondrial/synaptosomal membrane preparation of rat olfactory
bulb, and to study the distribution of CRF binding sites in discrete
regions of the rat CNS. The binding of 125I-Tyro rat/human CRF (125I-rCRF)
was time- and temperature-dependent, was sensitive to the pH, ionic
strength, and cationic composition of the incubation buffer, and was linear
over a broad range of membrane protein concentrations. 125I-rCRF binding to
olfactory bulb membrane was saturable, reversible, and, on Scatchard
analysis, revealed a high- affinity component with an apparent equilibrium
dissociation constant (Kd) of 0.2 nM and a low-affinity binding site with
Kd of approximately 20 nM. Data from pharmacological studies indicated that
the ability of a variety of CRF fragments and analogs to inhibit 125I-rCRF
to olfactory bulb membranes correlates well with their reported relative
potencies in stimulating pituitary adrenocorticotropic hormone secretion in
vitro. Consistent with a coupling of CRF receptors to adenylate cyclase,
the binding of 125I-rCRF was decreased by guanine nucleotides and increased
by magnesium ions. A heterogeneous distribution of 125I-rCRF binding sites
was found in the rat CNS, with highest densities present in olfactory bulb,
cerebellum, cerebral cortex and striatum, and progressively lower but
significant levels of binding were detected in cervical spinal cord,
hypothalamus, medulla, midbrain, thalamus, pons, and hippocampus. These
data, using a rat CRF ligand homologous to the endogenous peptide, are
consistent with those from previous studies demonstrating the presence of
specific binding sites for ovine CRF in rat brain, and provide further
support for the suggestion that endogenous CRF may function as a
neurotransmitter in the CNS.
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