Comparison of the contractile effects and binding kinetics of endothelin‐1 and sarafotoxin S6b in rat isolated renal artery

To date, only two mammalian endothelin (ET) receptors, termed ET A and ET B , have been cloned, sequenced and characterized. However, several functional studies of isolated blood vessels suggest that ET‐1‐induced contractions may be mediated by multiple ET A receptors. In this study, the ET A receptors in renal arteries isolated from Wistar rats were characterized by isometric tension recording and radioligand binding techniques. ET‐1, sarafotoxin S6b (StxS6b) and ET‐3 produced concentration‐dependent contraction with similar response maxima in endothelium‐denuded arteries, whereas the ET B receptor‐selective agonist StxS6c was inactive. ET‐1 and StxS6b were equipotent and 30 times more potent than ET‐3. This agonist profile, together with the findings that the ET A receptor‐selective antagonists, BQ‐123 and FR‐139317 caused concentration‐dependent, rightward shifts of the concentration‐effect curves to each agonist indicated that ET‐1‐induced contractions in rat renal artery were mediated via ET A receptors. BQ‐123 and FR‐139317 were both significantly more potent inhibitors of contractions induced by StxS6b or ET‐3 than of responses to ET‐1, raising the possibility that a component of ET‐1‐induced contraction was mediated through atypical, BQ‐123 (or FR‐139317)‐insensitive ET A receptors. However, in competition binding studies, specific [ 125 I]‐ET‐1 and [ 125 I]‐StxS6b binding to rat renal artery sections was completely abolished by BQ‐123 in a manner consistent with an action at a single site. Thus, competition binding studies did not provide any supportive evidence of the existence of a BQ‐123‐insensitive ET A receptor. Additional studies revealed marked differences in the kinetics of [ 125 I]‐ET‐1 and [ 125 I]‐StxS6b binding. Following a 3 h period of association of [ 125 I]‐ET‐1 with its receptors, no significant dissociation of receptor‐bound [ 125 I]‐ET‐1 was observed during a 4 h washout period. In stark contrast, dissociation studies revealed that specific [ 125 I]‐StxS6b binding to ET A receptors was reversible ( t 0.5diss , 100 min). A series of association binding studies were also consistent with the specific binding of [ 125 I]‐ET‐1 and [ 125 I]‐StxS6b being irreversible and reversible processes, respectively. Thus, differences in BQ‐123 potency against ET‐1 and StxS6b‐induced contractions in rat renal arteries might be due to differences in the kinetics of agonist binding, rather than due to the existence of atypical ET A receptors.British Journal of Pharmacology (1997) 121 , 253–263; doi: 10.1038/sj.bjp.0701126