Insulin-like growth factor binding protein 7, a member of insulin-like growth factor signal pathway, involved in immune response of small abalone Haliotis diversicolor

Abstract Insulin-like growth factor binding protein 7 (IGFBP7), the only member of the IGFBP superfamily that binds strongly to insulin, may have different functions from other IGFBPs. Unlike other IGFBPs, there is no knowledge available on aquatic invertebrate IGFBP7. In this study, a molluscan IGFBP7 gene, saIGFBP7 , was cloned for the first time from the small abalone Haliotis diversicolor . Its full-length cDNA sequence is 1812 bp, with a 720 bp open reading frame encoding a protein of 239 aa. The molecular mass of the deduced protein is approximately 25.37 kDa with an estimated pI of 5.00, and it shares highest 41% identity to IGFBP7 of Amblyomma americanum. Analysis of conserved domains revealed the presence of an IGFBP N-terminal domain (IB), a kazal-type serine proteinase inhibitor domain (KI), and an immunoglobulin-like C2 domain (IgC2) in saIGFBP7. Furthermore, the 12 cysteine residues and the signature amino acid motif ‘xCGCCxxC’ which are characterized by the amino terminus region of the IGFBP superfamily are all presented in saIGFBP7. Quantitative real-time PCR and western blot were employed to investigate the tissue distribution of saIGFBP7, and its expression under bacterial challenge. The saIGFBP7 mRNA and protein could be detected in all examined tissues, with the highest expression level in hemocytes, higher expression level in gills, and was up-regulated in hemocytes and gills after bacterial injection. In addition, saIGFBP7 mRNA transcripts were observed in a subset of the branchial epithelium and the nucleus of hemocytes using the in situ hybridization method. Interestingly, saIGFBP7 was detected mainly in the goblet-like cell of the branchial epithelium by immunohistochemistry. These results suggested that saIGFBP7 was likely to be involved in a function associated with pathogenic infection and may play an important role in the adult abalone immune system.