Characterization of a Y1-preferring NPY/PYY receptor in HT-29 cells.

Academic Article

Abstract

  • Equilibrium binding studies showed that butyrate-treated HT-29 cells express a high-affinity 125I-labeled peptide YY (125I-PYY) binding site with a dissociation constant of 0.32 +/- 0.12 nM (mean +/- SE, n = 4). This site was Y1 preferring because neuropeptide Y (NPY) and the Y1-selective agonist [Leu31,Pro34]NPY were equipotent to PYY at displacing 125I-PYY; PYY-(13-36) and pancreatic polypeptide were > 1,000- and 10,000-fold less potent at displacing the radioligand. PYY and [Leu31,Pro34]NPY inhibited forskolin-stimulated adenosine 3',5'-cyclic monophosphate production 63% and 48%, respectively, with a half-maximal inhibitory concentration between 0.1 and 1.0 nM. PYY and [Leu31,Pro34]NPY had no effect on release of intracellular calcium alone or on the increase in intracellular calcium concentration caused by carbachol or neurotensin. Northern blot analysis of poly(A)+ RNA from HT-29 cells demonstrated a single transcript of 2.5 kb that hybridized to a human Y1-receptor cDNA probe. Sequence analysis of a reverse transcription-polymerase chain reaction product amplified with primers based on human Y1-receptor cDNA confirmed that these cells contained mRNA encoding the human Y1 receptor. These studies show that butyrate-treated HT-29 cells constitutively express the Y1-preferring NPY/PYY receptor and Y1 mRNA and provide a new model for studies of PYY-regulated epithelial cell function and tissue-specific expression of the human Y1-receptor gene.
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    Keywords

  • Binding Sites, Blotting, Northern, Butyrates, Butyric Acid, Calcium, Cell Line, Cyclic AMP, Gastrointestinal Hormones, Humans, Intestinal Mucosa, Intracellular Membranes, Peptide YY, Peptides, Polymerase Chain Reaction, Receptors, Gastrointestinal Hormone, Receptors, Neuropeptide Y, Transcription, Genetic
  • Digital Object Identifier (doi)

    Author List

  • Mannon PJ; Mervin SJ; Sheriff-Carter KD
  • Start Page

  • G901
  • End Page

  • G907
  • Volume

  • 267
  • Issue

  • 5 Pt 1