EXAMINATION OF THE ROLE OF INHIBITION OF CYCLIC-AMP IN ALPHA(2)-ADRENOCEPTOR MEDIATED CONTRACTIONS OF THE PORCINE ISOLATED PALMAR LATERAL VEIN

1 We have examined the effect of elevation of cellular adenosine 3':5'-cyclic monophosphate (cyclic AMP) on alpha(1)- and alpha(2)-adrenoceptor-mediated contraction of the isolated palmar lateral vein of the pig. Cellular cyclic AMP was increased by either inhibition of phosphodiesterase by rolipram, or direct activation of adenylyl cyclase by forskolin. 2 Noradrenaline (1 nM-10 mu M) caused concentration-dependent contractions of the porcine isolated palmar lateral vein (pD(2) 7.32 +/- 0.07, n = 10). The selective alpha(1)-adrenoceptor antagonist, prazosin (0.1 mu M) and the selective alpha(2)-adrenoceptor antagonist, rauwolscine (1 mu M) caused a 10 fold rightward displacement of the concentration-response curve and a combination of the two antagonists caused a 200 fold rightward displacement of the concentration-response curve. The selective alpha(2)-adrenoceptor agonist, UK-14304, also produced concentration-dependent contractions of the palmar lateral vein (pD(2) 7.70 +/- 0.15, n = 5), but the maximum response was 55.5 +/- 7.6% (n = 5) of that produced by noradrenaline. Prazosin (0.1 mu M) failed to affect responses to UK-14304 but rauwolscine, 1 mu M, caused a 200 fold rightward displacement. The estimated pK(B) value for rauwolscine (8.28 +/- 0.19, n = 10) is consistent with inhibition of alpha(2)-adrenoceptors. Thus, the porcine isolated palmar lateral vein has a population of alpha(1)- and alpha(2)-adrenoceptors capable of producing a contraction. 3 Rolipram, 10 mu M, and forskolin, 1 mu M, caused a 2-3 fold rightward displacement of the noradrenaline concentration-response curve (CRC), but 1,9-dideoxyforskolin, 1 mu M, a forskolin analogue which does not activate adenylyl cyclase, failed to produce a significant inhibition of noradrenaline-induced contractions. The combination of forskolin (1 mu M) and rolipram (10 mu M) were additive, producing a 20 fold rightward displacement of the noradrenaline CRC. 4 Responses to noradrenaline were similarly affected by a combination of rolipram (10 mu M) and prazosin (0.1 mu M) (isolation of alpha(2)-adrenoceptors) and the combination of rolipram (10 mu M) and rauwolscine (1 mu M) (isolation of alpha(1)-adrenoceptors), resulting in a 100 fold rightward displacement of the noradrenaline CRC. Although forskolin inhibited both alpha(1)- and alpha(2)-adrenoceptor-mediated contractions, the effects produced were not similar. In particular, noradrenaline, 0.3-3 mu M, produced a significant contraction in the presence of forskolin (1 mu M) and prazosin (0.1 mu M) (an alpha(1)-adrenoceptor-mediated response) but not in the presence of forskolin (1 mu M) and rauwolscine (1 mu M) (an alpha(1)-adrenoceptor-mediated response). 5 Five minute exposure to either rolipram (10 mu M) or forskolin (1 mu M) elevated [H-3]-cyclic AMP of the porcine isolated palmar lateral vein by approximately 70% and 150-200%, respectively. Neither noradrenaline (1 nM-100 mu M) nor UK-14304 (1 nM-100 mu M) affected basal levels of [H-3]-cyclic AMP, but both produced a concentration-dependent inhibition of forskolin-stimulated [H-3]-cyciic AMP accumulation with a pK(i) of 7.43 +/- 0.1 (n = 3) and 7.97 +/- 0.18 (n = 3), respectively. The effect of noradrenaline against forskolin-stimulated [H-3]-cyclic AMP accumulation was reversed by rauwolscine (1 mu M) but not by prazosin (0.1 mu M). In contrast, alpha(2)-adrenoceptor activation did not affect rolipram-induced elevation of [H-3]-cyclic AMP. 6 These findings indicate that alpha(2)-adrenoceptor contractions of the porcine isolated palmar lateral vein are not produced by reduction in cellular cyclic AMP per se. It is proposed that this response involves a novel signal transduction mechanism. However, when cellular cyclic AMP has been elevated by agents that stimulate adenylyl cyclase, rather than through inhibition of phosphodiesterase, the ability of alpha(2)-adrenoceptors to inhibit cyclic AMP formation may be of functional importance in vascular smooth muscle.