Summary Adenosine has been shown to have negative inotropic, chronotropic and dromotropic effects on the heart. The pharmacological profiles of these effects suggest that they are mediated via R
i (A
1) adenosine receptors, but a direct demonstration of these receptors is still missing. In the present study we report direct labelling of these receptors with (-)N
6-[
125I]-p-hydroxyphenylisopropyladenosine ([
125I]HPIA). The radioligand bound in a saturable and reversible manner to a crude membrane preparation, the
B
max-value was 30.5 fmol/mg protein and the
K
D-value 1.1 nmol/l. A similar affinity of the ligand was obtained in kinetic and competition experiments. Competition experiments with a variety of adenosine analogues gave a pharmacological profile characteristic of R
i adenosine receptors with high affinities of N
6-substituted derivatives and a marked stereospecificity for N
6-phenylisopropyladenosine (PIA). Purification of the membrane preparation by density gradient centrifugation resulted in a 30-fold increase in the number of binding sites which was paralleled by a similar increase in the number of binding sites for [
3H]ouabain. Guanine nucleotides decreased binding of [
125I]HPIA in a dose-dependent manner, but the IC
50-values were considerably higher than those reported in other tissues. Finally, binding of [
125I]HPIA appeared to be entropy-driven which has been shown to be characteristic of agonist binding to R
i adenosine receptors. These results suggest the presence of R
i adenosine receptors in ventricular myocardium which may be responsible for the mediation of the effects of adenosine and its analogues.Abbreviations [
125I]HPIA
(-)N
6-[
125I]-p-hydroxyphenylisopropyladenosine
- (-)IHPIA
(-)N
6-iodo-p-hydroxyphenylisopropyladenosine
- (+)/(-)PIA
(+)/(-)N
6-phenylisopropyladenosine
- CHA
N
6-cyclohexyladenosine
- NECA
5-N-ethylcarboxamidoadenosine
- App(NH)p
5-adenylylimidodiphosphate
- Gpp(NH)p
5-guanylylimidodiphosphate
相似文献