Project description:Sexual maturation causes loss of fish muscle mass and deterioration of fillet quality attributes that prevent market purpose. We recently showed that fillet yield and flesh quality are recovered in female trout after spawning. To gain insights into the molecular mechanisms regulating flesh quality recovery we conducted a time-course analysis of large-scale gene expression in trout muscle from spawning to 33 weeks post-spawning. Using an Agilent-based microarray plate-form, we identified about 5500 unique genes that were differentially expressed during the post-spawning period. These genes segregated into four major clusters with distinct temporal expression profile and functional categories. A first cluster containing around 2000 genes down-regulated after 4 weeks after spawning was enriched in genes linked to mitochondrial ATP synthesis, fatty acid catabolism and proteolysis. A second cluster comprising about 1200 genes with early and transient induction between 2 to 8 weeks post-spawning was enriched in genes involved in RNA processing, translation, ribosome biogenesis and protein folding. A third cluster containing about 700 genes up-regulated from 4 weeks post-spawning onwards was enriched in genes encoding ribosomal subunits or regulating translation and cell cycle. At last a fourth cluster which contained about 1000 genes with late up-regulation from 13 weeks post- spawning onwards, was dominated by genes encoding myofibrillar proteins and matricial components and genes involved in glycolysis. On the whole our study indicates that muscle tissue restoration and flesh quality recovery occurring after spawning is mainly associated with anaerobic ATP production, muscle fibre hypertrophic growth, fatty acid metabolism and extracellular matrix remodelling. The generation of the first database of genes associated with post-spawning muscle recovery will allow future mechanistic studies and facilitate identification of genetic markers for fish muscle yield and quality traits.

Project description:BackgroundSexual maturation causes loss of fish muscle mass and deterioration of fillet quality attributes that prevent market success. We recently showed that fillet yield and flesh quality recover in female trout after spawning. To gain insight into the molecular mechanisms regulating flesh quality recovery, we used an Agilent-based microarray platform to conduct a large-scale time course analysis of gene expression in female trout white muscle from spawning to 33 weeks post-spawning.ResultsIn sharp contrast to the situation at spawning, muscle transcriptome of female trout at 33 weeks after spawning was highly similar to that of female trout of the same cohort that did not spawn, which is consistent with the post-spawning flesh quality recovery. Large-scale time course analysis of gene expression in trout muscle during flesh quality recovery following spawning led to the identification of approximately 3340 unique differentially expressed genes that segregated into four major clusters with distinct temporal expression profiles and functional categories. The first cluster contained approximately 1350 genes with high expression at spawning and downregulation after spawning and was enriched with genes linked to mitochondrial ATP synthesis, fatty acid catabolism and proteolysis. A second cluster of approximately 540 genes with transient upregulation 2 to 8 weeks after spawning was enriched with genes involved in transcription, RNA processing, translation, ribosome biogenesis and protein folding. A third cluster containing approximately 300 genes upregulated 4 to 13 weeks after spawning was enriched with genes encoding ribosomal subunits or regulating protein folding. Finally, a fourth cluster that contained approximately 940 genes with upregulation 8 to 24 weeks after spawning, was dominated by genes encoding myofibrillar proteins and extracellular matrix components and genes involved in glycolysis.ConclusionOverall, our study indicates that white muscle tissue restoration and flesh quality recovery after spawning are associated with transcriptional changes promoting anaerobic ATP production, muscle fibre hypertrophic growth and extracellular matrix remodelling. The generation of the first database of genes associated with post-spawning muscle recovery may provide insights into the molecular and cellular mechanisms controlling muscle yield and fillet quality in fish and provide a useful list of potential genetic markers for these traits.

Project description:We have constructed a rainbow trout high-density oligonucleotide microarray by using all the available tentative consensus (TC) sequences from the Rainbow Trout Gene Index database (The Computational Biology and Functional Genomics Lab., Dana Farber Cancer Institute and Harvard School of Public Health). The Rainbow Trout Gene Index integrates research data from all available international rainbow trout genomic research projects. The newly designed microarray incorporates 37,394 unique transcript-specific oligonucleotide probes, 60-mer long each. The microarray was printed according to our design by Agilent Technologies using the 4 X 44-design format and contains 1417 Agilent control spots. The performance of the new microarray platform was evaluated by analyzing gene expression associated with the rainbow trout vitellogenesis-induced muscle atrophy. These chips can be ordered from Agilent using design number 016320. This microarray is anticipated to open new avenues of research that will aid in the development of novel strategies to enhance growth efficiency and quality in salmonid species. Keywords: Development of an oligo-array for rainbow trout The performance of the new microarray platform was evaluated by analyzing transcriptome response associated with the rainbow trout vitellogenesis-induced muscle atrophy. Severe muscle deterioration accompanies the physiological responses of the energetic demands of the rainbow trout spawning/vitellogenesis. Atrophying muscle of fertile fish had 11% less extractable muscle (g/bw) and 11% less protein content compared to non-atrophying muscle of sterile fish (p<0.01). The rainbow trout was used to profile changes in gene expression of atrophying muscles. Gene expression levels were determined by comparing the amount of mRNA transcript present in the experimental sample (fertile fish) to the control (sterile fish). RNAs isolated from each experimental fish were run on separate microarrays in independent experiments, with no pooling. A total of 8 fish were used in the microarray experiments (4 replicates x 2 groups). Fluorophors (Cy3 and Cy5) were randomly assigned to RNA from each of the atrophying and nonatrophying muscles to limit the dye effect.

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Gene expression profiling of trout muscle during flesh quality recovery following spawning

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