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Transcriptome response to heat stress in the chicken hepatocellular carcinoma cell line

ABSTRACT: Background: Heat stress triggers an evolutionarily conserved set of responses in cells. The transcriptome responds to hyperthermia by altering expression of genes to adapt the cell or organism to survive the heat challenge. RNA-seq technology allows rapid identification of environmentally responsive genes on a large scale. In this study, we have used RNA -seq to identify heat stress responsive genes in the chicken male white-leghorn hepat ocellular (LMH) cell line. Result: The transcripts of 812 genes were responsive to heat stress (p <0.01) with 235 genes up- regulated and 577 down-regulated following 2.5 hours of heat stress. Among the up- regulated were genes whose products function as chaperones, along with genes aff ecting collagen synthesis and deposition, transcription factors, chromatin remodelers and genes modulating the WNT and TGF-beta pathways. Predominant among the down-regulated genes were ones that affect DNA replication and repair along with chromosom al segregation. Many of the genes identified in this study have not been previously implicated in the heat stress response. Conclusion: These data extend our understanding of the transcriptome response to heat stress. Many of the identified biological processes and pathways likely function in adapting cells and organisms to hyperthermic stress. This study may provide important guides to future efforts attempting to improve species abilities to withstand heat stress through genome wide association studies and breeding. In addition, the genes down regulated by heat stress may provide important targets for improving hyperthemic treatment in cancer patients.

ORGANISM(S): Gallus gallus

PROVIDER: GSE55321 | GEO | 2015/02/24

SECONDARY ACCESSION(S): PRJNA239321

REPOSITORIES: GEO

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Project description:Background In broilers, heat stress can result in reduced feed consumption, digestive inefficiency, impaired metabolism, and even death. The broiler sector of the U.S. poultry industry incurs approximately $52 million in heat stress-related losses annually. The objective of this study is to characterize the effects of chronic, cyclic heat stress on the transcriptome of a metabolically active organ, the liver. Characterizing the liver transcriptome of heat-stressed broilers will help clarify the effects of heat stress on metabolism. This information will provide a platform for future investigations that further elucidate physiologic responses to heat stress and seek methods to ameliorate the negative impacts of heat. Results Transcriptome sequencing of the livers of 8 broiler males using Illumina HiSeq 2000 technology, resulted in a total of 138 million, 100 base pair single end reads, yielding 13.8 gigabases of sequence. Forty genes were differentially expressed at a significance level of P-value < 0.05 and a fold change ≥ 2 in response to chronic, cyclic heat stress (mid-point of the last day of a 7-day cyclic heat stress of 7 hours per day), with 27 down-regulated and 13 up-regulated. Two gene networks were created from the function-based Ingenuity Pathway Analysis (IPA) of the differentially expressed genes; “Cell Signaling, Molecular Transport, Small Molecule Biochemistry” and “Endocrine System Development and Function, Small Molecule Biochemistry Cell Signaling”. Members of the MAPK signaling pathway and differentially expressed genes that are associated with MAPK-related functions were prominent in the networks. Cellular proliferation and differentiation, inflammationand stress-related signaling, and apoptosis-associated genes were down-regulated in response to heat stress. Genes responsible for inhibiting feed intake and sphingolipidrelated signaling were up-regulated. Genes involved with the regulation of inflammation, stress, thyroid hormone level, and body temperature were both up- and down-regulated. Conclusions Chronic, cyclic heat stress of broilers results in metabolic changes that can be characterized through RNA-seq analysis of the liver transcriptome. The primary affected pathways included cell signaling, molecular transport, endocrine system development and signaling, and small molecule biochemistry.

Project description:To gain insights into molecular mechanisms of tolerance to heat stress, we conducted a transcript profiling experiment to identify heat-responsive genes in contrasting peanut mini core accessions, either un-acclimated or acclimated to heat stress. Plants at reproductive stage were exposed to 28 °C (control), 45 °C for 15 d (un-acclimated) or 45 °C for 1 d followed by 7 d recovery and 15 d stress (acclimated). Two contrasting genotypes showing diverse response to stress were selected based on a bioassay involving chlorophyll fluorescence yield under elevated respiratory demand and membrane thermostability. Transcript profiling was performed using 4 x 44k custom oligo microarrays containing 22k peanut EST sequences. The microarray analysis identified 710 stress-induced and 770 stress-repressed putative heat-responsive transcripts in the tolerant genotype. Gene enrichment analysis was performed using Blast2GO program and genes with homology to known proteins were categorized into detailed molecular functional groups. Majority of stress-responsive genes assigned to KEGG pathways belonged to starch, sucrose and galactose metabolism followed by amino acid metabolism, and secondary metabolite biosynthesis. Differentially expressed transcripts from samples obtained from first year’s experiment were validated in the samples from second year by quantitative real-time PCR. Transcripts of eight genes involved in terpenoid and flavanoid biosynthesis were induced after second and seventh day, respectively, in leaves under heat stress. Metabolite analysis confirmed increases in metabolites of selected pathways under heat stress. The heat up-regulated genes in tolerant COC041 mini-core accession are potential candidate genes for engineering stress-tolerant peanuts and unraveling molecular mechanisms of peanut adaptation to heat stress. We used Agilent peanut microarrays to identify putative heat stress-responsive genes. Directly heat-stressed leaf tissues of the peanut genotypes COC041 (tolerant) and COC166 (susceptible) were used in the study. Three replications of microarray experiments were carried out by hybridizing the cRNA from different time points and stress conditions in a loop design on 4 x 44k microarray.

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Transcriptome response to heat stress in the chicken hepatocellular carcinoma cell line

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