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Rainbow trout red blood cells (RBCs): Control vs Heat shock, repeated samples from individuals

ABSTRACT: The retention of a nucleus in the mature state of fish red blood cells (RBCs), and the ability to easily collect and manipulate blood in non-terminal experiments, makes it an ideal tissue on which to study the cellular stress response in fish. Through the use of the cGRASP 16K salmonid microarray, we are currently investigating differences in RBC global gene transcription in fish held under control conditions (12C) and exposed to heat stress (one hour at 25C followed by recovery at 12C). Repeated blood sampling (via a dorsal aorta cannula) enables us to examine the individual stress response over time. Samples were taken pre-heat stress (representing individual control) and at 4 and 24 hours post heat stress (representing early and late transcriptional regulation). A total of ~3000 features had significant signal when hybridized with RBC RNA derived targets and cannulation did not have a significant effect on RBC mRNA expression at the investigated time points. Genes involved in the stress response, immune response, and apoptosis were among those showing the highest dysregulation during both early and late transcriptional regulation. Additionally, genes related to the differentiation and development of blood cells were upregulated at the twenty-four hour time point. This study enables a broader understanding of the mechanisms underpinning the stress response in fish and the discovery of novel genes that are regulated in a stress specific manner. Moreover, salmonid transcripts that are consistently dysregulated in blood in response to heat stress are potential candidates of non-lethal biomarkers of exposure to this particular stressor.

ORGANISM(S): Salmo salar Osmerus mordax Oncorhynchus tshawytscha Coregonus clupeaformis Oncorhynchus mykiss

PROVIDER: GSE21084 | GEO | 2010/09/21

SECONDARY ACCESSION(S): PRJNA126725

REPOSITORIES: GEO

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Project description:The retention of a nucleus in the mature state of fish red blood cells (RBCs), and the ability to easily collect and manipulate blood in non-terminal experiments, makes it an ideal tissue on which to study the cellular stress response in fish. Through the use of the cGRASP 16K salmonid microarray, we are currently investigating differences in RBC global gene transcription in fish held under control conditions (12C) and exposed to heat stress (one hour at 25C followed by recovery at 12C). Repeated blood sampling (via a dorsal aorta cannula) enables us to examine the individual stress response over time. Samples were taken pre-heat stress (representing individual control) and at 4 and 24 hours post heat stress (representing early and late transcriptional regulation). A total of ~3000 features had significant signal when hybridized with RBC RNA derived targets and cannulation did not have a significant effect on RBC mRNA expression at the investigated time points. Genes involved in the stress response, immune response, and apoptosis were among those showing the highest dysregulation during both early and late transcriptional regulation. Additionally, genes related to the differentiation and development of blood cells were upregulated at the twenty-four hour time point. This study enables a broader understanding of the mechanisms underpinning the stress response in fish and the discovery of novel genes that are regulated in a stress specific manner. Moreover, salmonid transcripts that are consistently dysregulated in blood in response to heat stress are potential candidates of non-lethal biomarkers of exposure to this particular stressor. Two condition experiment, control (11C) and heat stressed fish (one hour at 25C followed by return to 11C). Six fish in each treatment. 3 repeated samples taken from each individual at time 0, 4 hours post heat stress and 24 hours. Common reference design used, with all experimental samples (control or heat stress) combined with reference pool RBC RNA on each array.

Project description:MicroRNAs (miRNAs) are a class of small non-coding RNAs (18-25 nucleotides) that result in RNA silencing and post-transcriptional regulation of gene expression by binding to the 3'-untranslated regions of their target mRNAs. Mature miRNAs are widely reported in plants, animals and some viruses, which involved in the innate immune response, inflammatory diseases, cell signal transduction, and metabolism. Under temperature stress, miRNA expression is regulated to cope with the stress response. Up-regulation of 18 miRNAs and down-regulation of 11 miRNAs were found in the rat jejunum after heat treatment. 25 miRNA different expression under the cold and normal conditions were identified in zebrafish (Danio rerio). However, miRNAs involved in heat stress in GIFT have not been detected, and potentially important miRNA-signal pathway could have been overlooked. Study of the regulatory function of miRNA in heat-stress response is a crucial topic in the research of fish physiology. MiRNA expression profiles of tilapia in a variety of tissue have been studied recently by high-throughput next-generation sequencing technology (NGST). The expression levels of 218 miRNAs (e.g., miR-17, miR-29, miR-155 and miR-214) were significantly altered in tilapia kidney at 6-72h post- S. agalactiae infection. Additionally, 635 mature miRNAs were identified post-hatch day 5 in tilapia XX and XY gonads, in which 62 and 49 miRNAs showed different expression respectively (e.g., sex-biased miRNAs: miR-9, miR-21, miR-30a, miR-96, miR-200b, miR-212 and miR-7977). The gene expressions may contribute to immune regulation and stress adaptation in fish by heat stress. The levels of immune-related gene (e.g., immunoglobulin M, lysozyme, hepcidin and transferrin) expression were significantly up-regulated in turbot (Scophthalmus maximus) post-48h heat stress. HSPs play an important role in fish exposed to environmental stress. Some highly conserved HSPs could be synthesized in sculpins Oligocottus maculosus, turbot and GIFT in response to heat stress. Fatty acid composition and utilization in fish could be affected by heat stress. △6-desaturase-α and elongation of long chain fatty acids 5α (Elovl 5α) are involved in poly unsaturated fatty acid biosynthesis, which were obviously up-regulated in common carp (Cyprinus carpio) by heat stress, and help to increase fatty acid metabolism and maintain fluidity of cell membrane. However, in these studies the several genes were generated from known and functional protein-coding genes. The significance of mRNA transcriptome to heat-stress response in fish are far from clear. Thus, the study of miRNA and mRNA expression profiles may help us better understand the molecular mechanisms and signaling pathway of heat stress.

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Rainbow trout red blood cells (RBCs): Control vs Heat shock, repeated samples from individuals

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