Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiles in kidney of Japanese flounder injected with formalin-killed bacterial cells

ABSTRACT: In order to identify the genes induced by different bacterial cells, which may contain different types of pathogen associated molecular patterns (PAMPs), high-throughput gene expression analyses using Agilent custom-oligo DNA microarray containingon 9,573 probes constructed for Japanese flounder Paralichthys olivaceus were conducted. A number of genes showed significant changes in mRNA levels. However, there are no significant difference in a manner of the changes among the different bacterial cell treatments. The genes significantly induced by the treatments included well-known immune-related genes such as granulocyte-colony stimulation factor, haptoglobin, hepcidin. The kidney were isolated from the formalin killed cells (FKCs) intraperitoneal injected Japanese flounder, Paralichthys olivaceus using the four formalin-killed cells, Edwardsiella tarda strain 54, Lactococcus garviae strain EH8706, Streptococcus iniae strain 02 and Vibrio anguillarum strain H775-3, respectively. Fishes were administered by an intraperitoneal injection using the 1.0 x 10^7 to 1.0 x 10^8 cells of FKCs. After 6 hours from a injection, fish kidney was isolated. We also isolated phosphate-bufferd seline injected fish kidney as a control. We analyzed a four samples in control. We also analyze a three samples in FKC injected fish (16 hybridization).

ORGANISM(S): Paralichthys olivaceus

SUBMITTER: Hidehiro Kondo

PROVIDER: E-GEOD-29219 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The aquaculture industry has confronted severe economic losses due to infectious diseases in the last years. Piscirickettsiosis or Salmonid Rickettsial Septicaemia (SRS) is the bacterial disease caused by Piscirickettsia salmonis. This Gram-negative, non-motile, cellular pathogen has the ability to infect, survive, replicate, and propagate in salmonid monocytes/macrophages generating a systemic infection characterized by the colonization of several organs including kidney, liver, spleen, intestine, brain, ovary and gills. In this study, we attempted to determine whether global gene expression differences can be detected in different genetic groups of Atlantic salmon as a result of Piscirickettsia salmonis infection. Moreover, we sought to characterize the fish transcriptional response in order to reveal the mechanisms that might confer resistance in Atlantic salmon to an infection with Piscirickettsia salmonis. In doing so, after challenging with Piscirickettsia salmonis, we selected the families with the highest (HS) and the lowest (LS) recorded susceptibility for gene expression analysis using 32K cGRASP microarrays. Our results revealed in LS families expression changes are linked to iron depletion, as well as, low contents of iron in kidney cells and low bacterial load, indicated that the iron-withholding strategy of innate immunity is part of the mechanism of resistance against Piscirickettsia salmonis. This information contributes to elucidate the underlying mechanisms of resistance to Piscirickettsia salmonis infection in Atlantic salmon and to identify new candidate genes for selective breeding programmes. Forty full-sibling families of Atlantic salmon (Salmo salar) were infected by intraperitoneal injection with 0.2 mL Piscirickettsia salmonis (PS889, isolated from Oncorhynchus kisutch, 1M-CM-^W104 PFU/mL). After forty days, the fishes were harvested and the cumulative mortality (dead fish / total fish) for each family was calculated. For the second challenge, the six families with the highest cumulative mortality levels were considered of relatively high susceptibility (HS) and the six families with the lowest cumulative mortality levels were considered of relatively low susceptibility (LS) to the infection. Five control and five infected fish from three HS and three LS families were analyzed. For each HS and LS family, pools of RNA from control and infected fish were prepared separately and were reverse transcribed. Four slides were for each used family hybridized including two dye-swaped slides. Labeled samples were hybridized on a 32K cDNA microarray, developed at the Consortium for Genomics Research on All Salmonids Project (cGRASP), GEO accession number: GPL8904.

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Gene expression profiles in kidney of Japanese flounder injected with formalin-killed bacterial cells

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