Suppression subtractive hybridization coupled with microarray analysis to examine differential expression of genes in Japanese flounder Paralichthys olivaceus leucocytes during Edwardsiella tarda and viral hemorrhagic septicemia virus infection

Transcriptional changes in the peripheral blood leucocytes (PBL) of Japanese flounder Paralichthys olivaceus challenged by Edwardsiella tarda and viral hemorrhagic septicemia virus (VHSV) were investigated using suppression subtractive hybridization (SSH) coupled with cDNA microarray analysis. First...

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Bibliographic Details
Published in:Fish & shellfish immunology 2011-10, Vol.31 (4), p.524-532
Main Authors: Matsuyama, Tomomasa, Fujiwara, Atushi, Takano, Tomokazu, Nakayasu, Chihaya
Format: Article
Language:English
Subjects:
Animals
Edwardsiella tarda
Edwardsiella tarda - immunology
Enterobacteriaceae Infections - immunology
Enterobacteriaceae Infections - veterinary
Fish Diseases - immunology
Fish Diseases - microbiology
Fish Diseases - virology
Flounder
Gene Expression Regulation
Leukocytes - immunology
Leukocytes - metabolism
Marine
Microarray
Novirhabdovirus - immunology
Oligonucleotide Array Sequence Analysis - methods
Paralichthys olivaceus
Pleuronectiformes
Rhabdoviridae Infections - immunology
Rhabdoviridae Infections - microbiology
Rhabdoviridae Infections - veterinary
Suppression subtractive hybridization (SSH)
Viral hemorrhagic septicemia virus
Viral hemorrhagic septicemia virus (VHSV)
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Summary:Transcriptional changes in the peripheral blood leucocytes (PBL) of Japanese flounder Paralichthys olivaceus challenged by Edwardsiella tarda and viral hemorrhagic septicemia virus (VHSV) were investigated using suppression subtractive hybridization (SSH) coupled with cDNA microarray analysis. First, we constructed an SSH cDNA library using mRNA samples isolated from PBL of P. olivaceus that had been experimentally infected with E. tarda. We then examined the transcriptional changes occurring in the PBL due to E. tarda and VHSV infection using a cDNA microarray produced using clones produced from the SSH library. A total of 565 and 180 cDNA sequences corresponding to mRNA species that are either up- or down-regulated by E. tarda infection were isolated by SSH. While host gene expression responses in response to E. tarda and VHSV infection share several response elements, distinct patterns of gene expression were also observed. Specifically, E. tarda infection enhanced the expression of cell adhesion molecules while VHSV enhanced the expression of interferon and proteasome-related genes. In challenge trials of the two infectious agents, expression profiles of chemokines were also observed to differ. The results indicated that distinguishing between viral and bacterial infection is possible based on the RNA expression profiles of PBL from infected fish. ► We examined flounder leucocyte responses to the infection with E. tarda and VHSV. ► Distinct patterns of gene expression were observed between two pathogens. ► Virus and bacteria infection is distinguishable based on expression profiles.
ISSN:1050-4648
1095-9947
DOI:10.1016/j.fsi.2011.06.015