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Keywords:

Amino Acid Sequence, Animals, Base Sequence, Chickens, Cloning, Molecular, DNA, Complementary, Gene Expression Regulation, Hypothalamus, Molecular Sequence Data, Pituitary Gland, Anterior, Protein Isoforms, RNA, Messenger, Receptors, G-Protein-Coupled, Reverse Transcriptase Polymerase Chain Reaction, Sequence Alignment, Sequence Homology, Nucleic Acid, Tissue Distribution, Science & Technology, Life Sciences & Biomedicine, Endocrinology & Metabolism, Zoology, chicken, GHS-R, ghrelin, TRH, GH, GHRH, corticosterone, MESSENGER-RNA, RAT PITUITARY, GHRELIN, SECRETION, EXPRESSION, RELEASE, MECHANISMS, INVITRO, STOMACH, Receptors, Ghrelin, 0606 Physiology, 0608 Zoology, 0707 Veterinary Sciences, 3109 Zoology, 3202 Clinical sciences

Abstract:

Chicken ghrelin has recently been isolated as a hormone which stimulates growth hormone and corticosterone secretion in chicken. Ghrelin mediates these actions in mammals by binding to the growth hormone secretagogue receptor (GHS-R). In this study, we describe the partial cloning of two chicken GHS-R (cGHS-R) isoforms: cGHS-R1a and cGHS-R1c. cGHS-R1a and cGHS-R1c cDNA show, respectively, 81 and 78% homology with the corresponding parts of the human GHS-R1a cDNA. In contrast to the human GHS-R1b isoform, which is truncated after transmembrane domain 5 (TM-5), the chicken GHS-R1c isoform lacks 16 amino acids in TM-6 suggesting that this isoform is not active in ghrelin signal transduction. The cystein residues, N-linked glycosylation sites and potential phosphorylation sites, found in the human GHS-R1a, were also conserved in both chicken isoforms. RT-PCR analysis demonstrated cGHS-R1a and cGHS-R1c mRNA expression in all tissues tested, except liver and pancreas, with highest levels in the pituitary and the hypothalamus. Intermediate levels of expression were detected, in descending order, in the ovary, telencephalon, heart, adrenal gland, cerebellum, and optic lobes whereas low expression was detected in the brainstem, lung, kidney, proventriculus, duodenum, and colon. Very low expression was found in skin, stomach, and muscle. cGHS-R1c was expressed in lower amounts than cGHS-R1a in all analysed tissues. Administration of 1 microM chicken ghrelin to pituitaries in vitro resulted in a down-regulation of both cGHS-R isoforms within 15 min, whereas after 1h levels returned to control values. Growth hormone and corticosterone down-regulated cGHS-R1a and cGHS-R1c mRNA expression within 60 min of exposure, whereas growth hormone-releasing factor 1-29 (1 microM) only reduced cGHS-R1a mRNA expression after 60min. Thyrotropin-releasing hormone (1 microM) did not alter cGHS-R expression.