Molecular characterization of GPR84 in domestic cats

IF 2.1 3区医学 Q3 ENDOCRINOLOGY & METABOLISM General and comparative endocrinology Pub Date : 2024-04-18 DOI:10.1016/j.ygcen.2024.114520

Ichiro Yamamoto , Masaki Michishita , Koki Fujita , Tamami Sakai , Noriyasu Sasaki , Koh Kawasumi

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Abstract

G protein-coupled receptor 84 (GPR84) was cloned as an orphan receptor, and medium-chain fatty acids were then revealed as endogenous ligands. GPR84 is expressed in immune cells and is believed to protect liver function from lipotoxicity caused by overeating and high-fat diet intake. This study aimed to present the molecular characterization of GPR84 in domestic cats. The deduced amino acid sequence of the feline GPR84 shows high sequence homology (83–89 %) with the orthologues from other mammalians by cDNA cloning of feline GPR84. Remarkably high mRNA expression was observed in the bone marrow by Q-PCR analysis. The inhibition of intracellular cAMP concentration was observed in cells transfected with feline GPR84 and treated with medium-chain fatty acids. Immunostaining of GPR84 and free fatty acid receptor 2 (FFAR2)/GPR43 in the bone marrow, where high mRNA expression was observed, showed reactions in macrophages and myeloid cells. To clarify whether the receptor formed homo/hetero-merization, GPR84 and FFARs were analyzed using Nano-Luc binary technology and NanoLuc bioluminescence resonance energy transfer technologies, which revealed that GPR84 formed more heteromers with FFAR2 than homomers with each other. In addition, when GPR84 and FFAR2/GPR43 were cotransfected in the cell, their localization on the cell membrane was reduced compared with that when single receptors were transfected. These results indicated that GPR84 is a functional receptor protein that is expressed in cat tissues and may have a protein–protein interaction with FFAR2/GPR43 on the cell membrane.

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家猫 GPR84 的分子特征

G 蛋白偶联受体 84（GPR84）作为一种孤儿受体被克隆出来，随后中链脂肪酸作为内源性配体被揭示出来。GPR84 在免疫细胞中表达，被认为能保护肝脏功能免受暴饮暴食和高脂肪饮食引起的脂肪毒性的影响。本研究旨在介绍家猫 GPR84 的分子特征。通过克隆猫科动物 GPR84 的 cDNA，推导出的 GPR84 氨基酸序列与其他哺乳动物的同源物序列具有很高的同源性（83-89 %）。通过 Q-PCR 分析，在骨髓中观察到显著的高 mRNA 表达。在转染了猫科 GPR84 并用中链脂肪酸处理的细胞中，观察到细胞内 cAMP 浓度受到抑制。在骨髓中观察到高 mRNA 表达的 GPR84 和游离脂肪酸受体 2 (FFAR2)/GPR43 的免疫染色显示在巨噬细胞和骨髓细胞中出现反应。为明确受体是否形成同源/异源聚合，利用 Nano-Luc 二元技术和 NanoLuc 生物发光共振能量转移技术对 GPR84 和 FFARs 进行了分析，结果显示 GPR84 与 FFAR2 形成的异源聚合体多于相互之间形成的同源聚合体。此外，当 GPR84 和 FFAR2/GPR43 共转染到细胞中时，它们在细胞膜上的定位比单一受体转染时有所降低。这些结果表明，GPR84是一种在猫组织中表达的功能性受体蛋白，它可能与细胞膜上的FFAR2/GPR43发生蛋白-蛋白相互作用。

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来源期刊

General and comparative endocrinology 医学-内分泌学与代谢

CiteScore

5.60

自引率

7.40%

发文量

120

审稿时长

2 months

期刊介绍： General and Comparative Endocrinology publishes articles concerned with the many complexities of vertebrate and invertebrate endocrine systems at the sub-molecular, molecular, cellular and organismal levels of analysis.