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

• Three NCAM members exist in mandarin fish, and mfNCAM1b is fish specific.
• mfNCAMs express constitutively in both systemic and mucosal immune tissues.
• mfNCAM1a and mfNCAM1b can be induced by LPS or poly I:C though with different pattern.
• The lymphoid cells coexpressed mfNCAM1 and mfNCCRP-1 imply the equivalents to NK cells.

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.