Identification, expression analysis, and functional characterization of ghrelin and its receptors in spotted sea bass (
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RESEARCH PAPER
Identification, expression analysis, and functional characterization of ghrelin and its receptors in spotted sea bass (Lateolabrax maculatus) Peng Yu1 · Yangyang Zhou1 · Xin Qi1 · Hongying Fan1 · Kaiqiang Zhang1 · Xiaoyan Zhang1 · Yun Li1 · Haishen Wen1 Received: 19 January 2020 / Accepted: 15 May 2020 © Ocean University of China 2020
Abstract Ghrelin (Ghrl), an appetite-inducing peptide hormone secreted by the stomach, is the endogenous ligand for the growth hormone secretagogue receptor (Ghs-r). In this study, we identified the preproghrelin gene and its receptors in spotted sea bass (Lateolabrax maculatus). The ghrl gene consisted of an open reading frame (ORF) of 324 nucleotides encoding 107 amino acids, and the premature protein contained a 20-amino-acid mature peptide. Through a syntenic analysis, we also validated the annotation of growth hormone secretagogue receptor 1a (ghs-r1a) and growth hormone secretagogue receptor 1a-like (ghs-r1a-like), which contained seven-transmembrane structures, in spotted sea bass. The ORF of ghs-r1a consisted of 1152 bp that encoded a 383-amino-acid protein, and ghs-r1a-like contained an ORF of 2631 bp and produced a protein with 876 amino acids. A phylogenetic analysis showed that spotted sea bass ghrl and its receptors clustered with those of other fishes and were more distantly related to those of other vertebrates. In situ hybridization revealed that ghrl was highly expressed in the stomach and localized in the mucosa and submucosa. The expression of these genes varied during short-term starvation in a time-dependent manner. In vitro studies showed that after incubation with Ghrl for 3 h enhanced the expression of motilin (mln), gastrin (gas) and cholecystokinin (cck), but this effect vanished after 6 h of incubation. In summary, Ghrl and its receptors might play important roles in the regulation of food intake in spotted sea bass. Keywords Ghrl · Ghs-r · In vitro experiment · Localization · Spotted sea bass · Short-term starvation
Introduction Ghrelin (Ghrl) was first isolated from the stomachs of rats and humans and was identified as an endogenous ligand for the growth hormone secretagogue receptor (Ghs-r), which is a G-protein-coupled receptor with seven transmembrane domains (Kojima et al. 1999). Ghrl is encoded by the preproghrelin gene, which also encodes the 23-amino-acid peptide obestatin, a peptide that can inhibit food intake and Edited by Jiamei Li. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s42995-020-00055-x) contains supplementary material, which is available to authorized users. * Yun Li [email protected] * Haishen Wen [email protected] 1
Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China
counteract the effect of Ghrl (Zhang et al. 2005). In addition to mammals, Ghrl has also been identified in some fishes, including goldfish (Carassius auratus) (Unniappan et al. 2002), eel (Anguilla anguilla) (Kaiya et al. 2003b), channel
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