Transcriptome response following administration of a live bacterial vaccine in Atlantic salmon (Salmo salar).
Antibacterial responses have been studied in Atlantic salmon following an acute intra peritoneal injection of a genetically attenuated (aroA(-)) strain of Aeromonas salmonicida known to elicit protective immunity. Three tissues were studied for transcriptional changes, the liver, head kidney and the gill. RNA was collected from fish 6, 12, 24 and 48 h following infection or at the same time points from fish injected with PBS as non-infected control. PCR-select cDNA subtraction libraries were constructed from pooled 24 and 48 h post infection RNA to identify up-regulated mRNAs. One thousand four hundred and eighty six cDNA clones were sequenced from enriched cDNA libraries, of which 71% had significant homologies to known functional proteins. Many of these clones have previously been un-characterised in Atlantic salmon. A salmonid cDNA microarray was used to further analyse the gene expression profile as the library construction in itself does not answer the dynamics of the response. The greatest increase in expression identified in the array analysis was a liver antibacterial peptide, hepcidin that was increased 11-fold following the challenge. A panel of clones were chosen for semiquantitative reverse transcriptase PCR from all time points sampled. These results indicated there were both temporal differences and tissue differences in the transcriptional response to bacterial exposure, potentially of relevance to the establishment of protection.
Martin
S A M
SA
School of Biological Sciences, Zoology Department, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ, UK. sam.martin@abdn.ac.uk
Blaney
S C
SC
Houlihan
D F
DF
Secombes
C J
CJ
D048409:Aeromonas salmonicida / Q000276:immunology*; D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology*; D018076:DNA, Complementary / Q000235:genetics; D005786:Gene Expression Regulation; D015723:Gene Library; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020317:Salmo salar / Q000235:genetics* / Q000276:immunology* / Q000382:microbiology; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic / Q000235:genetics*; D014613:Vaccines, Attenuated / Q000276:immunology
D001428:Bacterial Vaccines; D018076:DNA, Complementary; D012333:RNA, Messenger; D014613:Vaccines, Attenuated