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

P. J. Mannon, S. J. Mervin, K. D. Sheriff-Carter

Research output: Contribution to journalArticle

23 Scopus citations

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 done 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.

Original languageEnglish (US)
Pages (from-to)G901-G907
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume267
Issue number5 30-5
DOIs
StatePublished - 1994

Keywords

  • Northern blot
  • adenylyl cyclase
  • polymerase chain reaction

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

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