Characterization of the receptors for chicken GHRH and GHRH-related peptides: identification of a novel receptor for GHRH and the receptor for GHRH-LP (PRP).

Growth hormone-releasing hormone and its structurally related peptides, GHRH-like peptide (GHRH-LP) (also called PRP), peptide histidine-isoleucine (PHI), vasoactive intestinal polypeptide (VIP), and pituitary adenylate cyclase-activating polypeptide (PACAP), have been reported to play important physiological roles in pituitary and extrapituitary tissues of vertebrates; however, little is known about the identity of these GHRH-related peptide receptors in birds. In this study, 6 receptors for GHRH and GHRH-related peptides (cGHRHR(1), cGHRHR(2), cGHRH-LPR, cPAC(1), cVPAC(1), and cVPAC(2)) were cloned from chicken brain or pituitary, and their functionalities were examined in Chinese hamster ovary (CHO) cells using a pGL3-CRE-luciferase reporter system. Results showed that: (1) all receptors are G protein-coupled receptors functionally coupled to the intracellular signaling pathway; (2) 2 GHRH receptors (cGHRHR(1) and cGHRHR(2)) were identified, and both receptors could be potently activated by cGHRH; (3) cGHRH-LP could activate its specific receptor cGHRH-LPR (cPRP-R), and it also activated cGHRHR(1) and cGHRHR(2); and (4) PACAP could potently activate its receptors cPAC(1), cVPAC(1) and cVPAC(2); however, cVPAC(1) and cVPAC(2) could also be effectively activated by cVIP and tPHI, indicating that they can serve as VIP receptors and potential PHI receptors. Using a reverse transcription polymerase chain reaction assay, we further examined the mRNA expression of these receptors in adult chicken tissues. The expressions of cGHRHR(1), cGHRHR(2), and cGHRH-LPR are restricted mainly to the pituitary and/or brain, whereas cPAC(1), cVPAC(1), and cVPAC(2) are expressed in most of the tissues examined. Collectively, our study identified the receptors for chicken GHRH and GHRH-related peptides, including a novel GHRH receptor (cGHRHR(2)), and established a basis to elucidate the roles of these peptides in target tissues.

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