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The interaction of MC3R and MC4R with MRAP2a in rainbow trout (Oncorhynchus mykiss)

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Abstract

Melanocortin 3 and 4 receptors are two important neural G protein-coupled receptors that regulate energy homeostasis in vertebrates. Melanocortin receptor accessory protein 2 (MRAP2) is also involved in the regulation of food intake and body weight as a variable regulator of melanocortin receptors. Rainbow trout (Oncorhynchus mykiss) is a valuable cold-water fish cultured worldwide. In the rainbow trout model, we cloned and identified mrap2a, a paralog of mrap2. Rainbow trout mrap2a consisted of a 690 bp ORF and was expected to encode a putative protein of 229 amino acids. The qPCR results showed that rainbow trout mrap2a was expressed at high levels in brain tissue similar to mc3r and mc4r. In addition, co-immunoprecipitation verified that MRAP2a interacts with MC3R and MC4R in vitro and that MRAP2a is involved in and regulates the constitutive activity and signaling of MC3R and MC4R. MRAP2a reduced constitutive and agonist-stimulated cAMP levels of MC3R; furthermore, MRAP2a increased constitutive ERK1/2 activation but reduced ligand-induced stimulation at high levels of expression. For MC4R, MRAP2a showed decreased cAMP basal activity but increased agonist-stimulated cAMP signaling and increased ACTH ligand sensitivity. However, MRAP2a failed to affect MC4R constitutive activity and agonist-induced ERK1/2 signaling. Undoubtedly, our study will have great significance for revealing the conserved role of MC4R and MC3R signaling in teleost fish, especially in cold-water fish growth and energy homeostasis.

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Data availability

The data that support the findings of this study are available from the corresponding author (Li-Xin Wang) upon reasonable request.

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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 and partially supported by the National Key R&D Program of China (2019YFD0900200).

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Authors and Affiliations

  1. College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Hui-Xia Yu, Yang Li, De-Bin Zhong, Hao-Lin Mo, Ze-Bin Jiang, Jia-Jia Yu, Dong-Mei Xiong, Hai-Xia Liu & Li-Xin Wang

  2. Meixian Aquaculture Farm of Shitouhe Reservoir Administration, Xianyang, 712000, Shaanxi, China

    Xin Ren

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  1. Hui-Xia Yu
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  2. Yang Li
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Contributions

Hui-Xia Yu: Dedicated to cell experiments and manuscript writing

Yang-Li: Manuscript writing

De-Bin Zhong: Gene cloning and vector construction

Xin-Ren: Provided the site and rainbow trout for this study

Hao-Lin Mo: Data analysis and graphing

Ze-Bin Jiang: Total RNA extraction and real-time PCR

Jia-Jia Yu, Dong-Mei Xiong, and Hai-Xia Liu: Provided experimental guidance

Corresponding author

Correspondence to Li-Xin Wang.

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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.

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Yu, HX., Li, Y., Zhong, DB. et al. The interaction of MC3R and MC4R with MRAP2a in rainbow trout (Oncorhynchus mykiss). Fish Physiol Biochem 49, 61–74 (2023). https://doi.org/10.1007/s10695-022-01159-0

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