Neuropeptide Y induced attenuation of catecholamine synthesis in the rat mesenteric arterial bed

The effect of neuropeptide Y (NPY) on the basal and nerve stimulation-induced increase in norepinephrine synthesis was studied in the isolated and perfused mesenteric arterial bed of the rat. Tyrosine hydroxylation, the rate-limiting step in catecholamine (CA) biosynthesis, was assessed by measuring the accumulation of DOPA in the perfusate/superfusate overflow after perfusion of the mesenteric arterial bed with the decarboxylase inhibitor m-hydroxybenzyl hydralazine (NSD-1015). Treatment with NDS-1015 resulted in a time-dependent increase in DOPA production and nerve stimulation (8 Hz, supramaximal voltage, 2 ms duration) increased DOPA production even further. NPY 1 to 100 nM was observed to produce a concentration-dependent attenuation in both the basal and nerve stimulation-induced increase in DOPA formation. To come to an understanding of the NPY receptor subtype mediating the inhibition of CA synthesis, the rank order of potency of a series of NPY analogs with varying selectivity for NPY receptor subtypes including intestinal polypeptide (PYY), PYY 13-36, Leu36 Pro34 NPY, human pancreatic polypeptide (h-PP), and rat pancreatic polypeptide (r-PP) were determined. In addition, the effect of various selective NPY antagonists on the inhibitory effect of NPY was also examined. These included the Y1 antagonist BIB03304, the Y2 antagonist BIIE0246, and the Y5 antagonist CGP71683. The IC50's for NPY, PYY, PYY13-36, Leu31 Pro34 NPY, and hPP in inhibiting CA synthesis were 5, 7, 15, 30, and 33 nM respectively. rPP failed to inhibit CA synthesis. All 3 of the NPY antagonists produced attenuation of the NPY-induced inhibition of CA synthesis, but it took a combination of all 3 to completely block the effect of a maximal inhibitory concentration of NPY. These results demonstrate that NPY inhibits CA synthesis in the perfused mesenteric arterial bed and can do so by activation of a variety of receptors including the Y1, Y2, and Y5.