Rat spinal cord alpha 2-adrenoceptors are of the alpha 2A-subtype: comparison with alpha 2A- and alpha 2B-adrenoceptors in rat spleen, cerebral cortex and kidney using 3H-RX821002 ligand binding.
Binding of the alpha 2-adrenoceptor antagonist radioligand 3H-RX821002 was investigated in membranes from rat spinal cord, spleen, cerebral cortex and kidney. The ligand was found to bind to saturable binding sites with apparent uniform affinities within each tissue. Seven compounds, some of which have previously been reported to be selective for either alpha 2A- or alpha 2B-adrenoceptors, were used in competition with 3H-RX821002. By using computer modelling, competition curves generated for three of these compounds (ARC 239, prazosin and oxymetazoline) could be resolved into two site fits in the kidney, Kd's of the drugs being compatible with the notion that these sites corresponded to alpha 2A- and alpha 2B-adrenoceptors. Moreover, rauwolscine and yohimbine were found to be about 14 and 9-fold selective for alpha 2B-adrenoceptors in the kidney. In all other tissues studied drug competition curves were uniphasic and computer modelled into one site fits, drug Kd's being well correlated to those for the alpha 2A-adrenoceptor. In rat spinal cord 26 further drugs, which showed wide variation in structure, were evaluated in competition with 3H-RX821002. Of these compounds, competion curves of the agonists UK-14,304, (-) and (+) adrenaline were modelled into two site fits whereas those of the remaining compounds could be modelled only into one site fits. Since the high affinity site for UK-14,304, (-) and (+) adrenaline was eliminated when EDTA, Gpp(NH)p and 140 mM NaCl was present in the assay the heterogeniety observed in spinal cord was considered to be due to formation of high and low affinity conformations of the alpha 2-adrenoceptor for agonists. It is concluded that 3H-RX821002 is useful to label both alpha 2A- and alpha 2B-adrenoceptors in the rat. Moreover, the binding sites labelled by 3H-RX821002 in the spinal cord appear to consist of a single population of alpha 2-adrenoceptors of the alpha 2A-type.
Department of Pharmacology, Umeå University, Sweden.
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S
Uhlén
J E
JE
Wikberg
D000317:Adrenergic alpha-Antagonists / Q000378:metabolism*; D000818:Animals; D002540:Cerebral Cortex / Q000378:metabolism; D003198:Computer Simulation; D004146:Dioxanes; D019329:Idazoxan / Q000031:analogs & derivatives; D007668:Kidney / Q000378:metabolism; D008024:Ligands; D008297:Male; D011869:Radioligand Assay; D051381:Rats; D011919:Rats, Inbred Strains; D011942:Receptors, Adrenergic, alpha / Q000378:metabolism*; D013116:Spinal Cord / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism
D000317:Adrenergic alpha-Antagonists; D004146:Dioxanes; D008024:Ligands; D011942:Receptors, Adrenergic, alpha; C058525:2-methoxyidazoxan; D019329:Idazoxan