Radioligand binding studies of alpha 1-adrenoceptor subtypes in rat heart.
1. In order to characterize the alpha 1-adrenoceptor subtypes mediating positive inotropic effects of adrenaline (in the presence of propranolol) in rat right ventricular strips and the Ca2+ sources used to elicit them, we have used radioligand binding to identify the alpha 1-adrenoceptor subtypes present in rat heart and the alpha 1-adrenoceptor affinity and subtype-selectivity of various pharmacological tools. 2. Amitryptiline, mianserin, trimipramine, oxaprotiline, clonidine, chloroethylclonidine, phenoxybenzamine, BE 2254 and 8-OH-DPAT competed for [3H]-prazosin binding in rat heart, vas deferens, liver, spleen, cerebral cortex and hippocampus but none of them displayed detectable alpha 1-adrenoceptor subtype-selectivity; nitrendipine did not compete for [3H]-prazosin binding in concentrations up to 5 mumol 1(-1). 3. The alpha 1 A-adrenoceptor-selective, 5-methyl-urapidil, (+)-niguldipine, and to a lesser extent (-)-niguldipine competed for [3H]-prazosin binding in rat heart, vas deferens, cerebral cortex and hippocampus with shallow and biphasic curves; analysis of these curves demonstrated that rat heart contains alpha 1A-and alpha 1B-adrenoceptors in a 20:80 ratio. 4. Treatment of rat right ventricular strips with 100 mumol l-1 chloroethylclonidine for 30 min at 30 degrees C followed by 60 min washout reduced the number of alpha 1-adrenoceptors, as assessed by [3H]-prazosin saturation experiments, by 74%. Treatment with 100 micromol l(-1) CdCl2 did not affect number or affinity of cardiac alpha 1-adrenoceptors and combined treatment with chlorethylclonidine and CdCl2 reduced alpha 1-adrenoceptor number by 90%. 5. Treatment of rat right ventricular strips with chloroethylclonidine steepened 5-methyl-urapidil competition curves and increased the relative contribution of alpha 1A-adrenoceptors from 26 to 89%. Treatment with CdCl2 did not affect 5-methyl-urapidil competition curves and combined treatment with chloroethylclonidineand CdCl2 increased the relative contribution of alpha lA-adrenoceptors to 66%.6. We conclude that rat heart contains alpha 1A- and alpha 1B-adrenoceptors in a 20:80% ratio. Treatment withchloroethylclonidine reduces alpha 1B-adrenoceptor number by 96% but has only minor effects on alpha 1A-adrenoceptor density. Treatment with CdCl2 does not affect the number of either alpha 1-adrenoceptor subtype.
Michel
M C
MC
Department of Medicine, University of Essen, Germany.
Hanft
G
G
Gross
G
G
D058668:Adrenergic alpha-1 Receptor Antagonists; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D002104:Cadmium / Q000494:pharmacology; D019256:Cadmium Chloride; D002118:Calcium / Q000378:metabolism; D002712:Chlorides / Q000494:pharmacology; D003000:Clonidine / Q000031:analogs & derivatives / Q000494:pharmacology; D004837:Epinephrine / Q000494:pharmacology; D006352:Heart Ventricles / Q000187:drug effects; D066298:In Vitro Techniques; D008297:Male; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D010879:Piperazines / Q000494:pharmacology; D011224:Prazosin / Q000493:pharmacokinetics; D011869:Radioligand Assay; D051381:Rats; D017208:Rats, Wistar; D018340:Receptors, Adrenergic, alpha-1 / Q000187:drug effects / Q000378:metabolism*
D058668:Adrenergic alpha-1 Receptor Antagonists; D000317:Adrenergic alpha-Antagonists; D002712:Chlorides; D010879:Piperazines; D018340:Receptors, Adrenergic, alpha-1; D002104:Cadmium; C057446:5-methylurapidil; C051731:chlorethylclonidine; D019256:Cadmium Chloride; D003000:Clonidine; D002118:Calcium; D011224:Prazosin; D004837:Epinephrine