RESUMO
Cloning with subsequent in vitro and in vivo characterization of vascular neuropeptide Y (NPY) receptor subtypes in porcine and canine peripheral tissues was performed. RT-PCR with Y1 and Y2 receptor-specific primers, indicated expression of Y1 receptors in both kidney and spleen of dog and pig, and expression of Y2 receptors in pig spleen. In pig kidney, expression of Y1 receptor mRNA was located to intrarenal arteries, as demonstrated with in situ hybridization using human probes. The cloned and sequenced canine Y1, porcine Y1 and Y2 receptors revealed high homologies to previously characterized mammalian NPY receptors. Membrane and autoradiographic receptor binding studies showed specific high-affinity binding sites for the purported Y1-selective radioligands 125I-[Leu31Pro34]peptide YY (PYY) and 3H-BIBP 3226 in dog spleen, and for the putative Y2-selective 125I-PYY(3-36) in dog and pig spleen. In the pig in vivo, [Leu31Pro34]PYY, administered i.v., evoked vasoconstriction in spleen and kidney, actions that were potently inhibited by the non-peptide Y receptor antagonist SR 120107A. In contrast, PYY(3-36) evoked vasoconstriction only in spleen and this effect was not influenced by SR 120107A. NPY evoked renal and splenic vasoconstriction in the dog in vivo, vascular responses that were inhibited by both BIBP 3226 and SR 120107A. Furthermore, the Y1 receptor agonist [Leu31Pro34]NPY also caused vasoconstriction in dog kidney and spleen, whereas the putative Y2 agonist N-acetyl[Leu28Leu31]NPY(24-36) evoked no such vascular responses. It is concluded that the pig spleen is likely to contain Y1 and Y2 receptors, both involved in splenic vasoconstriction. In contrast, the Y1 receptor seems to be the sole vascular NPY receptor subtype in pig kidney. Moreover, Y1 receptors predominate in dog spleen and kidney. Furthermore, the cloned canine Y1 receptor and the porcine Y1 and Y2 receptors show great homologies to, and possess ligand requirement profiles in accordance with, the human forms.
