Enhanced contractions of the porcine isolated ear artery by the alpha2-adrenoceptor agonist UK14304 are uncovered by pharmacological manipulation. As both neuropeptide Y (NPY) receptors and alpha2-adrenoceptors are negatively-coupled to adenylyl cyclase in this tissue, we determined whether NPY is also able to produce an enhanced contraction in the same tissue, under the same conditions. NPY (0.1 microM) produced a small contraction of porcine isolated ear arteries which was 5.1+/-0.8% of the response to 60 mM KCl (n = 14). An enhanced NPY response was uncovered if the tissue was pre-contracted with 0.1 microM U46619, and relaxed back to baseline with 1-2 microM forskolin before the addition of NPY (49.8+/-5.3%, n = 14). Forskolin (1 microM) stimulated cyclic AMP accumulation in porcine ear artery segments in the presence of 0.1 microM U46619 and 1 mM isobutylmethylxanthine (IBMX), NPY (0.1 microM) inhibited this response by 40%, but had no effect on basal levels of cyclic AMP. An enhanced response to 0.1 microM NPY was also obtained after pre-contraction with 0.1 microM U46619 and relaxation with either SNP (28.9+/-5.7%, n = 14), or dibutyryl cyclic AMP (21.2+/-4.6%, n = 14). This indicates that at least part of the enhanced response to NPY is independent of the agonist's ability to inhibit adenylyl cyclase. In conclusion, an enhanced contraction to NPY in the porcine isolated ear artery can be obtained by prior pharmacological manipulation. The enhanced responses are mediated through adenylyl cyclase-dependent and independent pathways similar to those reported for alpha2-adrenoceptors in this preparation.