Differential gene expression in breast muscle is similar in heat stressed layers and boilers at control temperatures
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ABSTRACT: In total 32 Male broiler and 32 male layer birds at the age of 6wk were subjected to two treatment: control (21C temperature and 50% RH) and heat-stress (32C temperature and 80% RH) in four climate chambers. After exposing the birds to said temperature and humidity the breast tissues samples were taken to get RNA and perform microarray experiment. RNA samples from two birds, from same treatment, same breed, and same chamber were pooled before hybridization.
Project description:Chronological age is one of the important factors influencing muscle development and meat quality in chickens. To evaluate the protein expression profiles during skeletal muscle development, we performed a tandem mass tag (TMT)-based quantitative proteomic strategy in pectoralis major (breast muscle) of Beijing-You chicken (BYC) at the age of 90, 120 and 150 days. A total of 1,413 proteins in chicken breast muscle and 197 of them were differentially expressed (fold change ≥ 1.2 or ≤ 0.8333 and p < 0.05). There were 110 up- and 71 down-regulated proteins in 120 d vs. 90 d group, 13 up- and 10 down-regulated proteins in 150 d vs. 120 d group. The proteomic profiles of BYC at 120 d were very similar to those at 150 d and highly different from those at 90 d, suggesting that 120 d might be an important chronological age for BYC. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these differentially expressed proteins were mainly involved in the pathway of glycolysis/gluconeogenesis, adrenergic signaling in cardiomyocytes, focal adhesion, oocyte meiosis and phagosome. Protein expression analysis indicated that the differences in muscle growth rate between ages were regulated by proteins such as LDHA and ENO3, whereas ATP2A1 and HSP70 were associated with water-holding capacity (WHC), and PPP1CB and COL1A2 were suggested to lie in the role of intramuscular fat (IMF) deposition. In addition, RACK1 was thought to be crucial for the sexual maturation during chicken development. Furthermore, some DEPs were quantified using parallel reaction monitoring (PRM) to validate the results from TMT analysis. Overall, the present work could strengthen our view of the temporal expression profile during development and identify novel biomarkers for genetic breeding of chickens.
Project description:For this study, thymic transcriptome responses to an acute heat stress and/or lipopolysaccharide (LPS) were investigated in a broiler line (heat and disease susceptible) and an inbred Fayoumi line (heat and disease resistant) of chickens. In a 2 x 2 design, 22 day-old birds were exposed to heat stress (35°C for 7 hours), lipopolysaccharide (100 µg/kg average body weight per line), or both stressors. Thermoneutral temperature (25°C) and phosphate buffered saline were used as the respective controls. Tissue samples were collected from the thymus and used to isolate high quality RNA. cDNA libraries (n = 31) were constructed and sequenced on the HiSeq 2500.
Project description:For this study, bursal transcriptome responses to an acute heat stress and/or lipopolysaccharide (LPS) were investigated in a broiler line (heat and disease susceptible) and an inbred Fayoumi line (heat and disease resistant) of chickens. In a 2 x 2 design, 22 day-old birds were exposed to heat stress (35°C for 7 hours), lipopolysaccharide (100 µg/kg average body weight per line), or both stressors. Thermoneutral temperature (25°C) and phosphate buffered saline were used as the respective controls. Tissue samples were collected from the bursa of Fabricius and used to isolate high quality RNA. cDNA libraries (n = 31) were constructed and sequenced (2 technical replicates per library; 62 total datasets) on the HiSeq 2500.
Project description:Transgenerational effects of early experience on behavioural, hormonal and gene expression responses to acute stress in the precocial chicken. The data for the offspring of the birds in this experiment is available in ArrayExpress with accession E-MTAB-925.
Project description:Transgenerational effects of early experience on behavioural, hormonal and gene expression responses to acute stress in the precocial chicken. The present accession presents the data from the adult chicken of the first (parental) generation.To access data for the young chicken of the first (parental) generation see accession E-MTAB-924. The male offspring of the birds of the first (parental) generation is also available in ArrayExpress with accession E-MTAB-925.
Project description:Transgenerational effects of early experience on behavioural, hormonal and gene expression responses to acute stress in the precocial chicken. The data for the parental stressed and control birds is available in ArrayExpress with accession E-MTAB-924, while this experiment only concerns their offspring that never been exposed to early life stress.
Project description:New experiences can trigger changes in gene expression in the brain. To understand this phenomenon better, we studied zebra finches hearing playbacks of birdsong. Earlier research had shown that initial playbacks of a novel song transiently increase the ZENK (ZIF-268, EGR1, NGFIA, KROX-24) mRNA in the auditory forebrain, but the response selectively habituates after repetition of the stimulus. Here, using DNA microarray analysis, we show that novel song exposure induces rapid changes in thousands of RNAs, with even more RNAs decreasing than increasing. Habituation training leads to the emer- gence of a different gene expression profile a day later, accompanied by loss of essentially all of the rapid "novel" molecular responses. The novel molecular profile is characterized by increases in genes involved in transcription and RNA processing and decreases in ion channels and putative noncoding RNAs. The M-bM-^@M-^XM-bM-^@M-^XhabituatedM-bM-^@M-^YM-bM-^@M-^Y profile is dominated by changes in genes for mitochondrial proteins. A parallel proteomic analysis [2-dimensional difference gel electrophoresis (2D-DIGE) and sequencing by mass spectrometry] also detected changes in mito- chondrial proteins, and direct enzyme assay demonstrated changes in both complexes I and IV in the habituated state. Thus a natural experience, in this case hearing the sound of birdsong, can lead to major shifts in energetics and macromolecular metabolism in higher centers in the brain. Adult male zebra finches were acoustically isolated and exposed to silence, novel song, or familiar song (exposure to testing song for 3 hours on the day prior to testing) on test day. The auditory lobule (AL) was collected 30 minutes after the onset of the testing experience. All samples were hybridized against the universal SoNG reference RNA pool, 6 biological replicates per group in each of 3 groups.
Project description:Songbirds provide rich natural models for studying the relationships of brain anatomy, behavioral function, environmental signals and gene expression. Under the Songbird Neurogenomics Initiative, investigators from more than a dozen laboratories collected brain samples from six species of songbird under a range of experimental conditions, and 488 of these samples were analyzed systematically for gene expression by microarray. ANOVA was used to test 32 planned contrasts in the data, revealing the relative impact of different factors. The brain region from which tissue was taken had the greatest influence on gene expression profile, affecting the majority of signals measured by the 18,848 non-redundant cDNAs spotted on the microarray. Social and environmental manipulations had highly variable impact, ranging from nil in some cases to robust in others. Several specific genes were identified as points of interest in the evolution of mechanisms by which environmental signals influence behavior. The data were also analyzed using Weighted Gene Co-expression Network Analysis (WGCNA) followed by Gene Ontology (GO) analysis. This revealed modules of co-regulated genes that are also enriched for specific functional annotations such as M-bM-^@M-^\ribosomeM-bM-^@M-^] (expressed more highly in juvenile brain) and M-bM-^@M-^\dopamine metabolic processM-bM-^@M-^] (expressed more highly in striatal song control nucleus Area X). These results underscore the complexity of influences on neural gene expression and provide a resource for studying how these influences are integrated during natural experience. RNA samples were collected from six different songbird species (phylogeny in SI Appendix, Figure S1) and representing 80 different M-bM-^@M-^\treatmentsM-bM-^@M-^], i.e., combinations of species, brain region, sex, age, and behavioral state. The tissue samples were organized around 15 standalone experiments (Table 1 and SI Appendix, Table S1), contributed by investigators in a dozen different laboratories but analyzed under uniform conditions in a single laboratory as originally planned under the Songbird Neurogenomics Initiative. Each sample was hybridized to a zebra finch cDNA array along with a universal reference (a pool of zebra finch telencephalic RNA). e11: Auditory lobules (AL) were collected from adult male zebra finches (Taeniopygia guttata) after 30 minutes of novel or familiar song playback, or silence control.
Project description:Stress in animals causes not only immediate reactions, but may modify many aspects of their biology for long periods. Particular interest has been paid to perinatal stress, and in several species this may affect behaviour and physiology for a long time, even across generations. Also adolescence has been shown to be a sensitive period in mammals, but so far, no systematic comparison has been performed of the relative sensitivity of different life phases. In this study, groups of chickens were exposed to a six-day period of repeated and varied stress during three different life phases: early (two weeks, 2W), adolescence (eight weeks, 8W) and adult (17 weeks, 17W), and the effects compared to an unstressed control group. Long-term and transgenerational effects were determined with hypothalamic gene expression analysis.
Project description:Photoperiod and hormonal cues drive dramatic seasonal changes in structure and function of the avian song control system. Little is known, however, about the patterns of gene expression associated with seasonal changes. Here we address this issue by exposing GambelM-bM-^@M-^Ys white-crowned sparrows to season-appropriate cues and extracting RNA from the telencephalic song control nuclei HVC and RA across multiple time points that capture different stages of growth and regression. We chose HVC and RA because while both nuclei change in volume across seasons, the cellular mechanisms underlying these changes differ. We thus hypothesized that different genes would be expressed between HVC and RA. We tested this by using the extracted RNA to perform a cDNA microarray hybridization developed by the SoNG initiative. We then validated these results using qRT-PCR. Supporting our hypothesis, only 59 of the 363 genes of interest were found to vary by more than |1.5| fold in expression in both nuclei, while 132 gene expression changes were HVC specific and 172 genes were RA specific. We then assigned many of these genes to functional categories relevant to the different mechanisms underlying seasonal change in HVC and RA, including neurogenesis, apoptosis, cell growth, dendrite arborization and axonal growth, angiogenesis, endocrinology, growth factors, and electrophysiology. This revealed categorical differences in the kinds of genes regulated in HVC and RA. These results show that different molecular programs underlie seasonal changes in HVC and RA, and that gene expression is time specific across different reproductive conditions. Our results provide insights into the complex molecular pathways that underlie adult neural plasticity. Experimental groups: long-term Short Day (SD); Long Day (LD) with Testosterone (T) for 3, 7, and 21 days (3LD+T, 7LD+T, 21LD+T respectively); SD and removal of T at 1 and 2 days (1SD-T, 2SD-T); two tissues (HVC, RA) collected separately from each individual animal; one experimental sample and one universal SoNG reference sample per array; dye balanced within groups. Six biological replicates per group, five biological replicates in 3LD+T.HVC group.