Abstract
Pinctada fucata martensii, is an economically important marine bivalve species cultured for seawater pearls. At present, we know little about the molecular mechanisms of the insulin signalling pathway in this oyster. Herein, we cloned and analysed an insulin-like peptide (PfILP) and its signalling pathway-related genes. We detected their expression levels in different tissues and developmental stages. Recombinant PfILP protein was produced and found to significantly increase primary mantle cell activity and induce the expression of the proliferating cell nuclear antigen (PCNA) gene. PfILP could also regulate the 293T cell cycle by stimulating the S phase and inhibiting the G1 and G2 phases. Recombinant PfILP protein induced the expression of its signalling pathway-related genes in mantle cells. In vitro co-immunoprecipitation analysis showed that PfILP interacts with PfIRR. PfILP activated expression of the pfIRR protein, and also activated the mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways by stimulating phosphorylation of MAPK and AKT. Further analysis showed that PfILP up-regulated glycogen synthesis-related genes glycogen synthase kinase-3 beta (GSK-3β), protein phosphatase 1 (PP1) and glucokinase (GK) at the mRNA level, as well as the expression of the PP1 protein, and phosphorylation of GSK-3β. These results confirmed the presence of a conserved insulin-like signalling pathway in pearl oyster that is involved in cell activity, glycogen metabolism, and other physiological processes.