Evidence for the existence of 'atypical' beta-adrenoceptors (beta 3-adrenoceptors) mediating relaxation in the rat distal colon in vitro.

1. Experiments were carried out to characterize the adrenoceptors mediating relaxant responses in the rat distal colon. Three agonists were used: noradrenaline, isoprenaline and the beta 3-adrenoceptor agonist BRL 37344. Phentolamine, propranolol and (+/- )-cyanopindolol were tested as antagonists. Tone in the rat distal colon was induced with KCl (30-40 mM) as a spasmogen, and relaxations of this KCl-induced tone produced by the agonists were measured. 2. Relaxant responses to noradrenaline that were obtained in the presence of propranolol (1 microM) were not antagonized by phentolamine (0.01 to 1 microM). Relaxant responses to isoprenaline that were obtained in the presence of phentolamine (1 microM) were antagonized in a concentration-dependent manner by propranolol (0.01 to 3 microM), although this antagonism was weak and non-competitive. Relaxant responses to BRL 37344 that were obtained in the presence of phentolamine (1 microM) were only weakly antagonized by high (1 microM) concentrations of propranolol. 3. Tachyphylaxis to BRL 37344 was observed, a second concentration-response curve being shifted to the right by 15 fold. Exposure of the tissues to BRL 37344 (1 microM) between concentration-response curves also caused rightward shifts in the responses to noradrenaline (18 fold) and isoprenaline (19 fold) but not to papaverine. 4. In the presence of phentolamine (1 microM) and propranolol (1 microM), the rank order of potency of the agonists was: (-)-isoprenaline (1.0) greater than or equal to BRL 37344 (0.93) greater than (-)-noradrenaline (0.3). 5. Responses to BRL 37344 in the presence of phentolamine (1 microM) and propranolol (1 microM) were antagonized by (+/-)-cyanopindolol (I microM), with an apparent pA2 value of 6.67. Responses to isoprenaline, under the same conditions, were antagonized in a competitive manner by (+/-)-cyanopindolol (0.1 to 10 microM), with the slope of the Schild plot close to unity and a pA2 value of 7.12. 6. The resistance of the relaxant responses to antagonism by phentolamine and propranolol, along with the relatively high potency of the beta 3-adrenoceptor agonist BRL 37344 and the antagonism of 'resistant' responses by (+/-)-cyanopindolol would suggest that 'atypical' beta 6-adrenoceptors, similar to the beta 3-adrenoceptors of rat adipocytes and other tissues, exist in the rat distal colon.