Characterization of glucagon-like peptide 2 receptor (GLP2R) gene in chickens: functional analysis, tissue distribution, and developmental expression profile of GLP2R in embryonic intestine

Domest Anim Endocrinol. 2014 Jul:48:1-6. doi: 10.1016/j.domaniend.2014.01.002. Epub 2014 Jan 18.

Abstract

This study characterized the glucagon-like peptide 2 receptor (GLP2R) gene of chickens because relatively little is known about the underlying mechanism of GLP2 actions in nonmammalian species. With the use of reverse transcription PCR, we first cloned the chicken GLP2R (cGLP2R) from adult intestine, which was predicted to encode a 529-amino acid receptor precursor. With the use of a pGL3-CRE luciferase reporter system, we demonstrated that cGLP2R expressed in Chinese hamster ovary cells could be potently activated by cGLP2 (half maximal effective concentration, 1.06 nM) but not by its structurally related peptides, including the newly identified glucagon-like peptide, indicating that cGLP2R is a functional receptor specific to cGLP2. Reverse transcription PCR assay revealed that cGLP2R mRNA was widely expressed in adult chicken tissues, including pancreas and various parts of the gastrointestinal tract. With the use of quantitative real-time reverse transcription PCR assays, we further investigated the mRNA expression of cGLP2R and its potential downstream mediators, epidermal growth factor receptor (EGFR) ligands (heparin-binding EGF-like growth factor, epiregulin, and amphiregulin), in the distal duodenum of developing embryos. The mRNA expression levels of GLP2R and EGFR ligands (heparin-binding EGF-like growth factor and amphiregulin) were shown to increase (P < 0.05 or 0.01) during the late embryonic stages (E16 and E20), implying a potential coordinated action of GLP2 and EGFR ligands on embryonic intestine development. Taken together, our findings not only establish a molecular basis to explore the physiological roles of GLP2 in birds, but they also provide comparative insights into the roles of GLP2R and its ligand in vertebrates, such as its roles in embryonic intestine development.

Keywords: Chicken; EGFR ligand; Embryonic intestine; GLP2; GLP2R.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CHO Cells
  • Chick Embryo / metabolism*
  • Cricetinae
  • Cricetulus
  • EGF Family of Proteins / genetics
  • EGF Family of Proteins / metabolism
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism
  • Gene Expression Regulation, Developmental / physiology*
  • Glucagon / metabolism
  • Intestinal Mucosa / metabolism
  • Intestines / embryology*
  • Molecular Sequence Data
  • Plasmids
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, Glucagon / genetics
  • Receptors, Glucagon / metabolism*
  • Transcriptome

Substances

  • EGF Family of Proteins
  • RNA, Messenger
  • Receptors, Glucagon
  • Glucagon
  • ErbB Receptors