Isolation of RAG-1 and IgM transcripts from the striped trumpeter (Latris lineata), and their expression as markers for development of the adaptive immune response

• Published in: Fish & shellfish immunology 2013-03, Vol.34 (3), p.778-788
• Main Authors: Covello, J.M., Bird, S., Morrison, R.N., Bridle, A.R., Battaglene, S.C., Secombes, C.J., Nowak, B.F.
• Format: Article
• Language: English
• Subjects: Adaptive Immunity; Amino Acid Sequence; Animals; Antibody; Biomarkers - metabolism; Cloning, Molecular; DNA, Complementary - analysis; DNA, Complementary - genetics; Fish Proteins - chemistry; Fish Proteins - genetics; Fish Proteins - metabolism; Gene expression; Gene Expression Regulation; Gene Expression Regulation, Developmental; Homeodomain Proteins - chemistry; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; IgM; Immunoglobulin M - chemistry; Immunoglobulin M - genetics; Immunoglobulin M - metabolism; Larva - genetics; Larva - growth & development; Larva - immunology; Larva - metabolism; Latris lineata; Maternal transfer; Molecular Sequence Data; Organ Specificity; Ovum - growth & development; Ovum - immunology; Ovum - metabolism; Perciformes - genetics; Perciformes - growth & development; Perciformes - immunology; Perciformes - metabolism; Phylogeny; RAG; Real-Time Polymerase Chain Reaction - veterinary; RNA, Messenger - analysis; RNA, Messenger - genetics; Sequence Alignment - veterinary; Sequence Analysis, DNA - veterinary; Tasmania
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2012.12.015

A partial sequence of the recombination activating gene-1 (RAG-1) and several full length sequences of the immunoglobulin M (IgM) heavy chain mRNA were obtained from the striped trumpeter (Latris lineata). The RAG-1 fragment consisted of 205 aa and fell within the core region of the open reading frame. The complete IgM heavy chain sequences translated into peptides ranging between 581 and 591 aa. Both genes showed good homology to other vertebrate sequences. The expression of the two genes was assessed throughout the early developmental stages of striped trumpeter larvae (5–100 dph) and used as markers to follow the ontogeny of the adaptive immune response. Using RT-PCR, RAG-1 mRNA expression was detectable at 5 dph and remained so until 80 dph, before becoming undetectable at 100 dph. IgM expression was also detectable at 5 dph, and remained so throughout. These patterns of expression may suggest that the striped trumpeter possess mature B cells with surface IgM at 100 dph. However, complete immunological competence is likely not reached until some time later. The early detection of IgM mRNA at 5 dph led to the investigation of its presence in oocytes. Both RAG-1 and IgM mRNA transcripts were detected in unfertilized oocytes, suggesting that they are maternally transferred. The biological significance of such a phenomenon remains to be investigated. ► Cloned and sequenced IgM C region and partial RAG-1 core region from Latris lineata. ► Both genes detected at 5 days post-hatch. ► RAG-1 expression ceased by 100 dph; mature B-cells after this time. ► mRNA from both genes found in unfertilized oocytes. ► Further investigation in maternal transfer of mRNA required.