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Thromb Haemost 1985; 54(02): 402-408
DOI: 10.1055/s-0038-1657749
Original Article
Schattauer GmbH Stuttgart

Studies of Alpha2-Adrenergic Receptors of Intact and Functional Washed Human Platelets by Binding of 3H-Dihydroergocryptine and 3H-Yohimbine-Correlation of 3H-Yohimbine Binding with the Potentiation by Adrenaline of ADP-Induced Aggregation

François Lanza
The Laboratoire de Biologie et de Pharmacologie des Interactions du Sang avec les Vaisseaux et les Biomatériaux, Centre Régional de Transfusion Sanguine, Strasbourg, France
,
Jean-Pierre Cazenave
The Laboratoire de Biologie et de Pharmacologie des Interactions du Sang avec les Vaisseaux et les Biomatériaux, Centre Régional de Transfusion Sanguine, Strasbourg, France
› Author Affiliations
Further Information

Publication History

Received 20 February 1985

Accepted 22 April 1985

Publication Date:
18 July 2018 (online)

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Summary

The binding of 3H-dihydroergocryptine and 3H-yohimbine to intact, discoid, functional, washed human platelets resuspended in Tyrode’s buffer containing Ca2+, Mg2+, human albumin and apyrase, was studied at 37° C. The binding of 3H-dihydroergocryptine was rapid, reversible and saturable (KD = 19.3 ± 4.2 nM, Bmax = 2590 ± 670 sites per platelet). The results were difficult to interpret because the bound ligand was not easily dissociated. In contrast, 3H-yohimbine bound in a rapid, reversible and saturable fashion to one class of sites (KD = 8.1 ± 1 nM, Bmax = 395 ± 35 sites/platelet) with the characteristics of alpha2-adrenergic receptors. Adrenaline alone did not aggregate intact platelets but potentiated ADP-induced aggregation. This effect of adrenaline was specifically inhibited by alpha2-antagonists such as yohimbine. The inhibition of 3H-yohimbine binding and the inhibition of the synergistic effect of adrenaline on ADP-induced aggregation by 16 different alpha- and beta-adrenergic compounds was significantly correlated (p <0.001). Thus, intact and functional washed human platelets can be used as a simple pharmacological model to screen alpha-adrenergic antagonists by measuring the inhibition of the potentiation of ADP-induced aggregation by adrenaline which is a direct reflection of the physiological effect of adrenaline on human platelet alpha2-adrenergic receptors. The inhibition constant derived from aggregation studies expresses the affinity of the ligand for its receptor as measured by more cumbersome binding studies with radioactive adrenergic antagonists such as 3H-yohimbine.

Keywords

Platelet - Alpha adrenergic receptor - Aggregation - Adrenaline - ADP - Yohimbine - Dihydroergocryptine
 

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