Abstract
The melanocortin-3 receptor (MC3R) is an important regulator of energy homeostasis and inflammation in mammals. However, its function in teleost fish needs to be further explored. In this study, we characterized rainbow trout MC3R (rtMC3R), which encoded a putative protein of 331 amino acids. Phylogenetic and chromosomal synteny analyses showed that rtMC3R was closely related to bony fishes. Quantitative PCR (qPCR) revealed that the transcripts of rtMC3R were highly expressed in the brain and muscle. The cellular function of rtMC3R was further verified by the signal-pathway-specific luciferase reporter assays. Four agonists such as α-MSH, β-MSH, ACTH (1–24), and NDP-MSH can active rtMC3R, increasing the production of intracellular cAMP and upregulating MAPK/ERK signals. Moreover, we found that rtMC3R stimulated with α-MSH and NDP-MSH can significantly inhibit the NF-κB signaling pathway. This research will be helpful for further studies on the function of MC3R in rainbow trout, especially the role of energy metabolism and immune regulation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The data and materials that support the findings of this study are available from the corresponding author upon reasonable request.
References
Balthasar N, Dalgaard L T, Lee C E, et al (2005) Divergence of melanocortin pathways in the control of food intake and energy expenditure[J]. Cell 123(3):493–505. https://doi.org/10.1126/science.1233000
Bustin SA, Benes V, Garson JA et al (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622. https://doi.org/10.1373/clinchem.2008.112797
Butler AA, Kesteson RA, Khong K, Cullen MJ, Pelleymounter MA, Dekoning J, Baetscher M, Cone RD (2000) A unique metalolic sysdrone causes obesity in the melanocortin-3 receptor-deficient mouse. Endocrinology 141(9)3518–3521. https://doi.org/10.1210/endo.141.9.7791
Cai M, Hruby VJ (2016) The melanocortin receptor system: a target for multiple degenerative diseases. Curr Protein Pept Sci 17:488–496. https://doi.org/10.2174/1389203717666160226145330
Chai B, Li J-Y, Zhang W et al (2007) Melanocortin-3 receptor activates MAP kinase via PI3 kinase. Regul Pept 139:115–121. https://doi.org/10.1016/j.regpep.2006.11.003
Dores RM, Baron AJ (2011) Evolution of POMC: origin, phylogeny, posttranslational processing, and the melanocortins. Ann N Y Acad Sci 1220:34–48. https://doi.org/10.1111/j.1749-6632.2010.05928.x
Dores RM, Londraville RL, Prokop J et al (2014) Molecular evolution of GPCRs: Melanocortin/melanocortin receptors. J Mol Endocrinol 52:T29-42. https://doi.org/10.1530/JME-14-0050
Fan ZC, Sartin JL, Tao YX (2008) Molecular cloning and pharmacological characterization of porcine melanocortin-3 receptor. J Endocrinol 196:139–148. https://doi.org/10.1677/JOE-07-0403
FAO (2020) The State of World Fisheries and Aquaculture 2020. Sustainability in action.
Gantz I, Fong TM (2003) The melanocortin system. Am J Physiol Metab 284:E468–E474. https://doi.org/10.1152/ajpendo.00434.2002
Gantz I, Konda Y, Tashiro T et al (1993) Molecular cloning of a novel melanocortin receptor. J Biol Chem 268:8246–8250. https://doi.org/10.1016/s0021-9258(18)53088-x
Getting SJ (2006) Targeting melanocortin receptors as potential novel therapeutics. Pharmacol Ther 111:1–15. https://doi.org/10.1016/j.pharmthera.2005.06.022
Girardet C, Butler AA (2014) Neural melanocortin receptors in obesity and related metabolic disorders. Biochim Biophys Acta - Mol Basis Dis 1842:482–494. https://doi.org/10.1016/j.bbadis.2013.05.004
Ho JCW, Jacobs T, Wang Y, Leung FC (2012) Identification and characterization of the chicken galanin receptor GalR2 and a novel GalR2-like receptor (GalR2-L). Gen Comp Endocrinol 179:305–312. https://doi.org/10.1016/j.ygcen.2012.09.005
Huang H, Tao Y-X (2014) Functions of the DRY motif and intracellular loop 2 of human melanocortin 3 receptor. J Mol Endocrinol 53:319–330. https://doi.org/10.1530/JME-14-0184
Ichiyama T, Campbell IL, Furukawa S et al (1999) Autocrine α-melanocyte-stimulating hormone inhibits NF-κB activation in human glioma. J Neurosci Res 58:684–689. https://doi.org/10.1002/(SICI)1097-4547(19991201)58:5%3c684::AID-JNR9%3e3.0.CO;2-%23
Klovins J, Haitina T, Fridmanis D et al (2004a) The melanocortin system in fugu: determination of POMC/AGRP/MCR gene repertoire and synteny, as well as pharmacology and anatomical distribution of the MCRs. Mol Biol Evol 21:563–579. https://doi.org/10.1093/molbev/msh050
Klovins J, Haitina T, Ringholm A et al (2004b) Cloning of two melanocortin (MC) receptors in spiny dogfish: MC3 receptor in cartilaginous fish shows high affinity to ACTH-derived peptides while it has lower preference to γ-MSH. Eur J Biochem 271:4320–4331. https://doi.org/10.1111/j.1432-1033.2004.04374.x
Lee EJ, Lee S, Jung J et al (2001) Differential regulation of cAMP-mediated gene transcription and ligand selectivity by MC3R and MC4R melanocortin receptors. Eur J Biochem 268:582–591. https://doi.org/10.1046/j.1432-1327.2001.01900.x
Li JT, Yang Z, Chen HP et al (2016) Molecular cloning, tissue distribution, and pharmacological characterization of melanocortin-4 receptor in spotted scat, Scatophagus argus. Gen Comp Endocrinol 230–231:143–152. https://doi.org/10.1016/j.ygcen.2016.04.010
Liao S, Chen K, Xi B et al (2019) Molecular cloning, characterization, and expression analysis of Megalobrama amblycephala melanocortin receptor 3 during fasting. J Fish Sci China 26:445–456. https://doi.org/10.3724/SP.J.1118.2019.18262
Lisak RP, Benjamins JA (2017) Melanocortins, melanocortin receptors and multiple sclerosis. Brain Sci 7:1–18. https://doi.org/10.3390/brainsci7080104
Logan DW, Bryson-Richardson RJ, Pagán KE et al (2003) The structure and evolution of the melanocortin and MCH receptors in fish and mammals. Genomics 81:184–191. https://doi.org/10.1016/S0888-7543(02)00037-X
Manna SK, Aggarwal BB (1998) α-Melanocyte-stimulating hormone inhibits the nuclear transcription factor NF-κB activation induced by various inflammatory agents. J Immunol 161:2873–2880. https://doi.org/10.1097/00005768-199905001-00697
Metherell LA, Chapple JP, Cooray S et al (2005) Mutations in MRAP, encoding a new interacting partner of the ACTH receptor, cause familial glucocorticoid deficiency type 2. Nat Genet 37:166–170. https://doi.org/10.1038/ng1501
Mitchell S, Vargas J, Hoffmann A (2016) Signaling via the NFκB system. Wires Syst Biol Med 8:227–241. https://doi.org/10.1002/wsbm.1331
Novoselova TV, Chan LF, Clark AJL (2018) Pathophysiology of melanocortin receptors and their accessory proteins. Best Pract Res Clin Endocrinol Metab 32:93–106. https://doi.org/10.1016/j.beem.2018.02.002
Patel H, Montero-Melendez T, Greco K, Perretti M (2011) Melanocortin receptors as novel effectors of macrophage responses in inflammation. Front Immunol 2:41. https://doi.org/10.3389/fimmu.2011.00041
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution 4(4):406–425
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. https://doi.org/10.1038/nprot.2008.73
Selz Y, Braasch I, Hoffmann C et al (2007) Evolution of melanocortin receptors in teleost fish: the melanocortin type 1 receptor. Gene 401:114–122. https://doi.org/10.1016/j.gene.2007.07.005
Takahashi A, Davis P, Reinick C et al (2016) Characterization of melanocortin receptors from stingray Dasyatis akajei, a cartilaginous fish. Gen Comp Endocrinol 232:115–124. https://doi.org/10.1016/j.ygcen.2016.03.030
Teare KA, Pearson RG, Shakesheff KM, Haycock JW (2004) Alpha-MSH inhibits inflammatory signalling in Schwann cells. NeuroReport 15:493–498. https://doi.org/10.1097/00001756-200403010-00022
Yang L, Tao Y (2017) Biased signaling at neural melanocortin receptors in regulation of energy homeostasis. Biochim Biophys Acta - Mol Basis Dis 1863:2486–2495. https://doi.org/10.1016/j.bbadis.2017.04.010
Yang L, Zhang Z, Wen H, Tao Y (2019) Characterization of channel catfish (Ictalurus punctatus) melanocortin-3 receptor reveals a potential network in regulation of energy homeostasis. Gen Comp Endocrinol 277:90–103. https://doi.org/10.1016/j.ygcen.2019.03.011
Yang Y, Harmon CM (2017) Molecular signatures of human melanocortin receptors for ligand binding and signaling. Biochim Biophys Acta (BBA)-Molecular Basis Dis 1863:2436–2447. https://doi.org/10.1016/j.bbadis.2017.04.025
Yang Z, Huang Z, Tao Y (2015) Functions of DPLIY motif and helix 8 of human melanocortin-3 receptor. J Mol Endocrinol 55:107. https://doi.org/10.1530/JME-15-0116
Acknowledgements
We are grateful for the methodological guidance by Prof. Ya-Xiong Tao at Auburn University.
Funding
This research was supported by the National Natural Science Foundation of China (No. 31502180), Shaanxi Provincial Water Conservancy Science and Technology Fund (No. 2016slkj-16), and Shaanxi Provincial Agricultural Products Quality and Safety Project, partially supported by the National Key R&D Program of China (2019YFD0900200).
Author information
Authors and Affiliations
Contributions
Li-Xin Wang conceived and designed this study; Hui-Xia Yu and Yang Li cloned rainbow trout mc3r, performed the cellular experiment, and prepared the manuscript; Wei-Jia Song, Hui Wang, and Hao-Lin Mo collected the samples; Qiao Liu, Xin-Miao Zhang, and Ze-Bin Jiang extracted RNA and examined the tissue distribution of rtMC3R.
Corresponding author
Ethics declarations
Ethics approval
The Faculty Animal Policy and Welfare Committee of Northwest A&F University under contract (NWAFU-314020038) approved this experiment, and the experimental process complied with protocols of international guidelines for the ethical use of animals in research.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Yu, HX., Li, Y., Song, WJ. et al. Functional characterization of melanocortin-3 receptor in rainbow trout (Oncorhynchus mykiss). Fish Physiol Biochem 48, 241–252 (2022). https://doi.org/10.1007/s10695-021-01033-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-021-01033-5