TY - JOUR T1 - The Peptide YY-Preferring Receptor Mediating Inhibition of Small Intestinal Secretion Is a Peripheral Y<sub>2</sub> Receptor: Pharmacological Evidence and Molecular Cloning JF - Molecular Pharmacology JO - Mol Pharmacol SP - 124 LP - 134 DO - 10.1124/mol.60.1.124 VL - 60 IS - 1 AU - Mathieu Goumain AU - Thierry Voisin AU - Anne-Marie Lorinet AU - Robert Ducroc AU - Annick Tsocas AU - Claude Rozé AU - Patricia Rouet-Benzineb AU - Herbert Herzog AU - Ambikaipakan Balasubramaniam AU - Marc Laburthe Y1 - 2001/07/01 UR - http://molpharm.aspetjournals.org/content/60/1/124.abstract N2 - A peptide YY (PYY)-preferring receptor [PYY &gt; neuropeptide Y (NPY)] was previously characterized in rat small intestinal crypt cells, where it mediates inhibition of fluid secretion. Here, we investigated the possible status of this receptor as a peripheral Y2 receptor in rats. Typical Y2 agonists (PYY3–36, NPY3–36, NPY13–36, C2-NPY) and very short PYY analogs (N-α-Ac-PYY22–36 andN-α-Ac-PYY25–36) acting at the intestinal PYY receptor were tested for their ability to inhibit the binding of125I-PYY to membranes of rat intestinal crypt cells and of CHO cells stably transfected with the rat hippocampal Y2 receptor cDNA. Similar PYY preference was observed and all analogs exhibited comparable high affinity in both binding assays. The same held true for the specific Y2 antagonist BIIE0246 with a K i value of 6.5 and 9.0 nM, respectively. BIIE0246 completely abolished the inhibition of cAMP production by PYY in crypt cells and transfected CHO cells. Moreover, the antagonist 1) considerably reversed the PYY-induced reduction of short-circuit current in rat jejunum mucosa in Ussing chamber and 2) completely abolished the antisecretory action of PYY on vasoactive intestinal peptide (VIP)-induced fluid secretion in rat jejunum in vivo. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) experiments showed that Y2 receptor transcripts were present in intestinal crypt cells (3 × 102molecules/100 ng RNAT) with no expression in villus cells, in complete agreement with the exclusive binding of PYY in crypt cells. Finally, a full-length Y2 receptor was cloned by RT-PCR from rat intestinal crypt cells and also from human small intestine. We conclude that the so-called PYY-preferring receptor mediating inhibition of intestinal secretion is a peripheral Y2receptor. PYYpeptide YYNPYneuropeptide YPPpancreatic polypeptideCHOChinese hamster ovarypporcinerratVIPvasoactive intestinal peptideKRBKrebs-Ringer bufferRT-PCRreverse transcription-polymerase chain reactionTBETris-borate EDTAbpbase pair(s)TTXtetrodotoxinBIIE0246(S)-N2-[[1-[2-[4-[(R,S)-5,11-dihydro-6(6h)-oxodibenz[b,e]azepin-11-yl]-1-piperazinyl]-2-oxoethyl] cyclopentyl] acetyl]-N-[2-[1,2-di-hydro-3,5-(4H)-dioxo-1,2-diphenyl-3H-1,2,4-triazol-4-yl]ethyl]-argininamid ER -