Typical and atypical NK1 tachykinin receptor characteristics in the rabbit isolated iris sphincter

1. A contraction of the rabbit isolated iris sphincter smooth muscle follows activation of either tachykinin NK1 or NK3 receptors. We have here characterized the pharmacological activity profiles of various tachykinin receptor agonists considered to have NK1-receptor-preferring activity in this preparation. 2. Two groups of NK1-receptor-preferring agonists could be distinguished in terms of a common pharmacological profile. The first group (Group 1) included [Glp6,L-Pro9]-SP(6-11) (septide), [Glp6]-SP (6-11), substance P methyl ester, delta-aminovaleryl-[L-Pro9, N-MeLeu10]-SP(7-11) (GR73632), and [Apa9-10]-SP. The second group (Group 2) included [Pro9]-SP, substance P, physalaemin and ranamargarin. 3. Under control conditions, the responses to Group 1 agonists were relatively fast in offset (time for reversal of maximal responses, 11.2-18.2 min), and were antagonized by NK1-receptor-selective antagonists (range of pKB estimates vs various agonists; GR82334, 7.1-8.2; (+/-)-CP-96,345, 8.9-9.5; RP67580, 7.0-7.4). Following incubation of the tissue with phenoxybenzamine (20 microM, 10 min), the affinity of GR82334, tested against the Group 1 agonists, substance P methyl ester and septide, was not significantly different (P < 0.05; n = 7-18) to that determined in untreated tissues (substance P methyl ester pKB 7.5 +/- 0.1 and 7.2 +/- 0.2, respectively; septide 7.7 +/- 0.2 and 7.9 +/- 0.2, respectively). Further, response offset times (5.0-8.5 min) were little reduced as compared to those observed in untreated tissues. 4. Under control conditions, the response to Group 2 agonists was markedly slow in offset (times for reversal of maximal responses, 51.4-70.4min), and was not attenuated significantly by the NK1-receptor-selective antagonists GR82334 (I MicroM), (+/-)-CP-96,345 (0.1 MicroM) or RP67580 (1 MicroM). In contrast,after phenoxybenzamine pretreatment, responses to Group 2 agonists reversed rapidly (times for reversal of maximal responses, 13.1-24.2 min), and were now antagonized by GR82334 (pKB estimates, 6.4-7.1).5. The responses to the NK3-receptor-selective agonist Succ-[Asp6,Me-Phe8]-SP(6-l1) (senktide) were relatively fast in offset (time for reversal of maximal response was 18.6 +/- 1.7 min) and were not inhibited by GR82334 (10 MicroM; n = 5). The contractile response resulting from co-application of the Group 1 agonist, septide together with senktide, did not exhibit prolonged response offset kinetics.6. Assuming simple competition at equilibrium, these data from the rabbit iris smooth muscle could be explained either by interaction of the various ligands with two separately-existing NK1 receptor-subtypes or -isoforms; or alternatively by a preferential interaction of the two agonist groups with different binding domains on a common NK1 receptor.