Regional differences of the inhibition of GABAB ligand binding by the GTP analogue Gpp(NH)p.
In order to determine whether the interactions between GABAB receptors and G-proteins differ in several brain areas, we have used the reduction in high-affinity GABAB binding by the GTP analogue Gpp(NH)p as an internal assay marker for G-protein linkage to GABAB receptors. The results indicate that Gpp(NH)p inhibits the binding of the GABAB receptor agonist [3H]CGP 27492 (80 to 95%) in a biphasic manner between 0.1 nM and 1 mM. The IC50 for high-affinity sites is significantly higher in cerebellum (70 nM, 53% of binding sites) than in cortex, hippocampus, corpus striatum and thalamus (15-30 nM, 63-73% of binding sites). The IC50S of the low-affinity sites in hippocampus and cortex (170 microM and 210 microM, respectively) were significantly higher than the IC50S in cerebellum, thalamus and corpus striatum (18-39 microM). All these binding sites are sensitive to pertussis toxin (PTX; 7-15 micrograms/mg protein), implicating that they are linked either to Gi or to Gzero proteins. The two binding sites observed (high affinity, nM and low affinity, microM for Gpp(NH)p) and the regional dependence in affinity of these sites may originate either from different GABAB receptor subtypes, different G-proteins or different coupling mechanisms between G-proteins and GABAB receptors. Whereas the PTX site of G-protein linked to GABAB receptors changes with age [24], the GTP binding site does not differ between peripubertal rats (5-6 weeks) and adults rats (10-12 weeks).
Unité INSERM U.398, Faculté de Médecine, Strasbourg, France.
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Mathivet
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Bernasconi
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H
Bittiger
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Marescaux
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D002531:Cerebellum / Q000378:metabolism; D002540:Cerebral Cortex / Q000378:metabolism; D003342:Corpus Striatum / Q000378:metabolism; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism; D008297:Male; D037342:Pertussis Toxin; D011869:Radioligand Assay; D051381:Rats; D017208:Rats, Wistar; D018080:Receptors, GABA-B / Q000378:metabolism*; D013788:Thalamus / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D018080:Receptors, GABA-B; D010566:Virulence Factors, Bordetella; D006165:Guanylyl Imidodiphosphate; D037342:Pertussis Toxin