Evaluating the interactions between red stingray (Dasyatis akajei) melanocortin receptors and elephant shark (Callorhinchus milii) MRAP1 and MRAP2 following stimulation with either stingray ACTH(1-24) or stingray Des-Acetyl-αMSH: A pharmacological study in Chinese Hamster Ovary cells

• Published in: General and comparative endocrinology Vol. 265; pp. 133 - 140
• Main Authors: Dores, Robert M., Scuba-Gray, Michelle, McNally, Bridgette, Davis, Perry, Takahashi, Akiyoshi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2018
• Subjects: Callorhinchus milii; Dasyatis akajei; Elephant shark; Melanocortin receptors; MRAP1; MRAP2; Red stingray; Callorhinchus milii; Melanocortin receptors; Dasyatis akajei; Red stingray; Elephant shark; MRAP2; MRAP1
• ISSN: 0016-6480; 1095-6840
• DOI: 10.1016/j.ygcen.2018.02.018

•Co-expression with esMRAP1 increased sensitivity for srACTH(1-24) for all five stingray MCRs.•Co-expression with esMRAP1 did not increase sensitivity for Des-Acetyl-αMSH for any stingray MCR.•Co-expression with esMRAP2 only increased sensitivity for Des-Acetyl-αMSH for stingray MC4R.•Co-expression with esMRAP2 had not effect on sensitivity for srACTH(1-24) for any stingray MCR. Previous studies on bony vertebrate MC2R orthologs (i.e., ray finned fishes, amphibians, reptiles, birds, and mammals) have shown that these MC2R orthologs have an obligatory requirement for interaction with bony vertebrate MRAP1 orthologs to a) allow for the trafficking of the MC2R ortholog to the plasma membrane; and b) to allow activation by ACTH, but not by any MSH-sized ligand. In addition, previous studies have found that co-expression of teleost and mammalian MC4R orthologs with corresponding MRAP2 has positive effects on sensitivity to stimulation by αMSH or ACTH. MRAP1 and MRAP2 paralogs have been detected in the genome of a cartilaginous fish (elephant shark), yet two cartilaginous fish MC2R orthologs (elephant shark and red stingray) do not apparently require MRAP1 for trafficking to the plasma membrane when expressed in Chinese Hamster Ovary (CHO) cells, and both orthologs can be activated by either ACTH or MSH-sized ligands. This study was done to determine whether sensitivity to stimulation by ACTH(1-24) or Des-Acetyl-αMSH is affected when stingray (sr) MC1R, MC2R, MC3R, MC4R or MC5R were co-expressed in CHO cells with either elephant shark (es) MRAP1 or esMRAP2. The results indicated that co-expression with heterologous MRAP1 increased the sensitivity of all five stingray melanocortin receptors for srACTH(1-24), but had not statistically significant effect on stimulation by srDes-Acetyl-αMSH for any of the stingray melanocortin receptors. Conversely, co-expression with esMRAP2 only enhanced sensitivity for srDes-Acetyl-αMSH for srMC4R, but had no effect on the other stingray orthologs, and there was no increase in sensitivity for srACTH(1-24) for any of the stingray melanocortin receptors. It appears then that some stingray melanocortin receptors have retained the ability to interact with a cartilaginous MRAP1 paralog. These results are discussed with reference to radiation of MRAP-related accessory proteins in cartilaginous fishes.