ABSTRACT: De Novo assembly and transcriptome analysis of Atlantic salmon macrophage/dendritic-like TO cells following type I IFN treatment and Salmonid alphavirus subtype-3 infection
Project description:We used RNA-seq to characterize the transcriptomic changes induced by type I IFN treatment and salmon alphavirus subtype 3 (SAV-3) infection in TO-cells, a macrophage/dendritic like cell-line derived from Atlantic salmon (Salmo salar L) head kidney leukocytes.
Project description:Atlantic salmon (Salmo salar L.) is an environmentally and economically important organism and its gene content is reasonably well characterized. From a transcriptional standpoint, it is important to characterize the normal changes in gene expression over the course of early development, from fertilization through to the parr stage.S. salar samples were taken at 17 time points from 2 to 89 days post fertilization. Total RNA was extracted and cRNA was synthesized and hybridized to a new 44K oligo salmonid microarray platform. Quantified results were subjected to preliminary data analysis and submitted to NCBI’s Gene Expression Omnibus. Throughout the entire period of development, several thousand genes were found to be differentially regulated. This work represents the trancriptional characterization of a very large geneset that will be extremely valuable in further examination of the transcriptional changes in Atlantic salmon during the first few months of development. The expression profiles can help to annotate salmon genes in addition to being used as references against any number of experimental variables that developing salmonids might be subjected to.
Project description:Atlantic salmon (Salmo salar L.) is an environmentally and economically important organism and its gene content is reasonably well characterized. From a transcriptional standpoint, it is important to characterize the normal changes in gene expression over the course of early development, from fertilization through to the parr stage.S. salar samples were taken at 17 time points from 2 to 89 days post fertilization. Total RNA was extracted and cRNA was synthesized and hybridized to a new 44K oligo salmonid microarray platform. Quantified results were subjected to preliminary data analysis and submitted to NCBI’s Gene Expression Omnibus. Throughout the entire period of development, several thousand genes were found to be differentially regulated. This work represents the trancriptional characterization of a very large geneset that will be extremely valuable in further examination of the transcriptional changes in Atlantic salmon during the first few months of development. The expression profiles can help to annotate salmon genes in addition to being used as references against any number of experimental variables that developing salmonids might be subjected to. 17 time points (2 - 89 dpf) with three samples for each time point Equimolar amounts from all three individuals in each time point were pooled. Then reference pool was created by including equal quantities cRNA from each time point, except for the first time point which had less material included.
Project description:Salmon pancreas disease, caused by salmonid alphavirus (SAV), is an economically important disease affecting farmed Atlantic salmon (Salmo salar L.) in Europe, with outbreaks reported in Scotland, Norway, and Ireland. A microarray-based study was performed to evaluate the host transcriptomic response during the early stages of an experimentally induced salmonid alphavirus 1 (SAV 1) infection in Atlantic salmon (Salmo salar L.) Head kidney was sampled from five fish PD infected Atlantic salmon parr and uninfected controls on days 1, 3 and 5 post injection (d.p.i). RNA from tissue samples was amplified and interrogated using the 17k TRAITS / SGP cDNA microarray, with results validated by SYBR green real-time PCR. The greatest number of significantly differentially expressed genes was recorded on day 3 p.i. These were found to be mainly associated with immune and defence mechanisms including genes involved in interferon I & II pathways and major histocompatibility complex class I & II responses. The expression of genes associated with apoptosis (BcL2 and caspase 3/7) and cellular stress (heat shock protein) were also found to differ significantly between infected and uninfected individuals as were genes involved in inhibiting viral attachment and replication, such as ubiquitin, serum myeloid, and and viperin.
Project description:Piscirickettsiosis or salmonid rickettsial septicemia (SRS) caused by intracellular Gram-negative bacterial pathogen, Piscirickettsia salmonis, constitutes one of the main infectious diseases in salmonid aquaculture. In the present study, we aimed to explore the transcriptomic responses in the Atlantic salmon kidney following a low dose of EM-90-like P. salmonis infection using the consortium for Genomic Research on All Salmonids Project (cGRASP)-designed Agilent 44K salmonid oligonucleotide microarray. Two infection level phenotypes [low (L-SRS) and high (H-SRS) infection level] were revealed in infected individuals at 21 days post-injection (DPI) based on multivariate analyses of expression of four antibacterial biomarker transcripts (campb, hampa, il8a, tlr5a) and pathogen level measured by qPCR. Five fish from each group (Control, L-SRS, and H-SRS) were selected for transcriptome profiling. We identified 1636 and 3076 differentially expressed probes (DEPs) in the L-SRS and H-SRS groups, respectively (FDR = 0.01).
Project description:Gills of teleost fish represent a vital multifunctional organ; however, they are subjected to environmental stressors, causing gill damage. Gill damage is associated with significant losses in the Atlantic salmon aquaculture industry. Gill disorders due to environmental stressors are exacerbated by global environmental changes, especially with open-net pen aquaculture (as farmed fish lack the ability to escape those events). The local and systemic response to gill damage, concurrent with several environmental insults, are not well investigated. We performed field sampling to collect gill and liver tissue after several environmental insults. Using a 44K salmonid microarray platform, we aimed to compare the transcriptomes of pristine and moderately damaged gill tissue. The gill damage-associated biomarker genes and associated qPCR assays arising from this study will be valuable in future research aimed at developing therapeutic diets to improve farmed salmon gill health.