Identification,expression analysis,and functional characterization of ghrelin and its receptors in spotted sea bass(Lateolabrax maculatus)

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 la(ghs-rla)and growth hormone secretagogue receptor la-1ike(ghs-rla-like),which contained seven-transmembrane structures,in spotted sea bass.The ORF of ghs-rla consisted of 1152 bp that encoded a 383-amino-acid protein,and(ghs-rla-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.