Molecular cloning, tissue distribution, and pharmacological characterization of blunt snout bream (Megalobrama amblycephala) melanocortin-5 receptor
The melanocortin-5 receptor (MC5R) plays an important role in the regulation of exocrine secretion in mammals. Its function in fish is not well established. In this study, we reported the molecular cloning, tissue expression, and pharmacological characterization of Megalobrama amblycephala MC5R (MamMC5R), as well as the effect of catching stress on its expression. The full-length cDNA of Mammc5r gene was 1237 bp, consisted of a 990-bp open reading frame encoding 329 amino acids. Sequence analyses revealed that the nucleotide and amino acid sequences of Mammc5r were highly homologous (> 90%) with MC5Rs of zebrafish, common carp, and goldfish. Tissue expression profile analysis showed that Mammc5r was widely expressed in both central and peripheral tissues, with the highest expression in the brain. Catching stress significantly changed the expression of Mammc5r in the skin, brain, and eye. In the skin, the expression level of Mammc5r was significantly reduced at 1 h and 4 h and increased at 24 h after catching stress. The Mammc5r expression level was rapidly upregulated in the brain and eye at 1 h and then decreased to the level before stress at 4 h and 24 h. With human MC5R (HsaMC5R) as a control, several agonists including α-melanocyte-stimulating hormone (α-MSH), and β-MSH in addition to an analogue [Nle4, D-Phe7]-α-MSH (NDP-MSH), were used to investigate the binding and signaling properties of MamMC5R. The results revealed that MamMC5R had the highest affinity for NDP-MSH, followed by α- and β-MSH. Taken together, the data suggested that MamMC5R might play a role in stress response in M. amblycephala.
KeywordsMegalobrama amblycephala MC5R Gene cloning Tissue expression Pharmacological characterization
This work was supported by the Natural Sciences Foundation of China (No.31572662), and the earmarked fund for China Agriculture Research System (CARS-45), and the Auburn University Intramural Grant Program and Animal Health and Disease Research Program of College of Veterinary Medicine at Auburn University (to Y.-X. T).
Compliance with ethical standards
Animal experiments were approved by the Ethical Committee of our Center and followed the national guidelines for the care and use of vertebrate animals.
Conflict of interest
The authors declare that they have no conflict of interest.
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