Investigating the potential immune role of fish NCAMs: Molecular cloning and expression analysis in mandarin fish

Fish Shellfish Immunol. 2015 Oct;46(2):765-77. doi: 10.1016/j.fsi.2015.08.006. Epub 2015 Aug 12.

Abstract

The immune role of NCAMs has been revealed in mammals, yet there is no such report in fish. Hence, we analyzed the molecular characterizations and immune-associated expression patterns of NCAMs in mandarin fish. Three NCAM members, named mfNCAM1a, mfNCAM1b and mfNCAM2, were identified. Among the cDNA sequences of mfNCAMs, AU-rich elements in the 3' UTRs of mfNCAM1b and mfNCAM2 as well as VASE sequences in the fourth Ig-like domain-encoding regions of mfNCAM1a and mfNCAM1b were discovered. Moreover, the syntenic analysis suggested that the duplication of NCAM1 is fish-specific. At mRNA and protein levels, the expression analyses revealed that mfNCAMs existed in both systemic and mucosal immune tissues, and located within lymphoid cells. Upon stimulated either by LPS or poly I:C, the expression level of mfNCAM1a was significantly up-regulated in head kidney, spleen, liver, and gut, whereas mfNCAM1b only in head kidney and liver, and mfNCAM2 only in liver. Additionally, the cells coexpressed mfNCAM1 and mfNCCRP-1 might imply the equivalents to mammalian NK cells. Our finding firstly demonstrates the member-specific immune-related tissue expression pattern and immune activity for fish NCAMs. Current data indicate that mfNCAM2 has little immune activity, while the immune activity of mfNCAM1a exists in more tissues than mfNCAM1b, and mfNCAM1a may tend to respond more actively to viral while mfNCAM1b to bacterial stimulants. Additionally, NCAM1b should be a fish-specific member with unique immune function, judging from its different expression pattern, immune activity as well as phylogenetic relationship to mfNCAM1a.

Keywords: Mandarin fish; Mucosal immune; NCAM; NCCRP-1; NK cell; Systemic immune.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Immunohistochemistry / veterinary
  • In Situ Hybridization / veterinary
  • Lipopolysaccharides / pharmacology
  • Molecular Sequence Data
  • Neural Cell Adhesion Molecules / chemistry
  • Neural Cell Adhesion Molecules / genetics*
  • Neural Cell Adhesion Molecules / metabolism
  • Organ Specificity
  • Perciformes / genetics*
  • Perciformes / immunology*
  • Phylogeny
  • Poly I-C / pharmacology
  • Real-Time Polymerase Chain Reaction / veterinary
  • Sequence Alignment / veterinary
  • Up-Regulation

Substances

  • DNA, Complementary
  • Fish Proteins
  • Lipopolysaccharides
  • Neural Cell Adhesion Molecules
  • Poly I-C