title: Characterization of glucagon-like peptide 1 receptor (GLP1R) gene in chickens: functional analysis, tissue distribution, and identification of its transcript variants.
abstract: Glucagon-like peptide 1 (GLP1) receptor plays a critical role in mediating the biological actions of GLP1 in mammals and fish; however, the gene structure, expression, and functionality of GLP1 receptor (GLP1R) remain largely unknown in birds. In this study, the full-length cDNA of chicken GLP1R (cGLP1R) was first cloned from brain tissue by reverse transcription PCR. The putative cGLP1R is 459 amino acids in length and shares high amino acid sequence identity with that of human (79%), rat (80%), and Xenopus (75%). Using a pGL3-CRE luciferase reporter system, we found that cGLP1R expressed in Chinese hamster ovary cells could be potently activated by cGLP1 (EC(50), 0.11 nM) but not by other structurally related peptides, indicating that cGLP1R is a functional receptor specific to cGLP1. Interestingly, in addition to identification of the transcript encoding cGLP1R of 459 amino acids, eight transcript variants, which were generated by alternative mRNA splicing and predicted to encode either C-terminally or N-terminally truncated cGLP1Rs, were also identified from chicken brain or testis. In line with this finding, multiple cGLP1R transcripts were detected to be expressed in most chicken tissues examined, including pancreas, gastrointestinal tract, and various brain regions by reverse transcription PCR. Using the dual-luciferase reporter assay system, we further found that the 5'-flanking region of cGLP1R gene displays promoter activities in cultured HepG2 and HEK293 cells, suggesting that it may control cGLP1R gene transcription in chicken tissues, including nonpancreatic tissues. Taken together, the results from the present study establish a molecular basis to investigate the roles of GLP1 in chickens.
authors.affiliation: Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610064, China.
authors.forename: G
authors.initials: G
authors.lastname: Huang
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: H
authors.initials: H
authors.lastname: Fu
authors.forename: Z
authors.initials: Z
authors.lastname: Yan
authors.forename: G
authors.initials: G
authors.lastname: Bu
authors.forename: X
authors.initials: X
authors.lastname: He
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D002645:Chickens / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D052216:Glucagon-Like Peptide 1 / Q000235:genetics / Q000378:metabolism; D000067757:Glucagon-Like Peptide-1 Receptor; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018027:Receptors, Glucagon / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D018076:DNA, Complementary; C000593833:GLP1R protein, human; C000593835:Glp1r protein, rat; D000067757:Glucagon-Like Peptide-1 Receptor; D012333:RNA, Messenger; D018027:Receptors, Glucagon; D052216:Glucagon-Like Peptide 1