RNA-Seq of chicken cecal tonsil samples treated with chronic heat stress
Ontology highlight
ABSTRACT: To study the effect of chronic heat stress on mature laying type hens, RNA-Seq was performed on cecal tonsil samples during a 4-week heat stress experiment. RNA-Seq samples were collected 3-hour, 2-weeks, and 4-weeks post heat from both control (unheated) and treatment (heated) birds. RNA was extracted from the tissues with RNAqueous Total RNA Isolation Kit, treated with DNase, and mRNA cDNA libary were prepared with Illumina TrueSeq Stranded mRNA Library Prep Kit. Sequencing was done on 2 lanes with 12 libraries per lane on Illumina HiSeq 3000 platform.
Project description:To study the effect of chronic heat stress on producing laying type hens, RNA-Seq was performed on breast samples during a 4-week heat stress experiment. RNA-Seq samples were collected 3-hour, 2-weeks, and 4-weeks post heat from both control (unheated) and treatment (heated) birds. RNA was extracted from the tissues with RNAqueous Total RNA Isolation Kit, treated with DNase, and mRNA cDNA libary were prepared with Illumina TrueSeq Stranded mRNA Library Prep Kit. Sequencing was done on 4 lanes with 12 libraries per lane on Illumina HiSeq 3000 platform.
Project description:To study the effect of chronic heat stress on producing laying type hens, RNA-Seq was performed on liver samples during a 4-week heat stress experiment. RNA-Seq samples were collected 3-hour, 2-weeks, and 4-weeks post heat from both control (unheated) and treatment (heated) birds. RNA was extracted from the tissues with mirVana miRNA Isolation Kit, treated with DNase, and mRNA cDNA libary were prepared with Illumina TrueSeq Stranded mRNA Library Prep Kit. Sequencing was done on 4 lanes with 12 libraries per lane on Illumina HiSeq 3000 platform.
Project description:Calcium (Ca) and phosphorus (P) are essential micronutrients that are linked to a wide set of biological processes. In laying hens there is still uncertainty about the optimal Ca/P ratio in feed and further strategies for the reliable mineral restriction in poultry diets are required. The dataset is based on Lohmann Selected Leghorn (LSL) and Lohmann Brown (LB) laying hens sampled at peak performance. The experimental design comprises four dietary groups differed in Ca (recommended vs. 15 % reduction) and mineral P (adequate vs. 20 % reduction) levels; 1) control diet (Con; Ca=34.4g/kg, P=5.3 g/kg and Ca/P ratio=7.45), 2) Low Ca and P diet (LCaP), 3) low Ca diet (LCa), and low P diet (LP). Jejunal transcriptome profiles were assessed by mRNA sequencing in a total of 80 animals (10 hens per experimental group for each laying line) at sampling age of 31 weeks) to approximate the consequences of variable Ca and P supply.
Project description:Emerging data demonstrated that the gut microbiota plays an important role in protecting the integrity of the epithelial barrier, forming a mucosal immune system, and maintaining intestinal homeostasis through its metabolites. However, the intestinal microbiota community can be affected by environmental factors, such as litter, photoperiod, or temperature. Thus, we investigated the effect of different monochromatic light combinations on cecal microbiota composition as well as explored the molecular mechanism by how the external light color information mediate cecal tonsil T lymphocyte proliferation. In this study, a total of 160 chicks were exposed to monochromatic light [red (R), green (G), blue (B), or white (W) light] or green and blue monochromatic light combination (G→B) from P0 to P42. The 16S rRNA microbial sequencing results showed that the richness and diversity of the cecum microbiota and the abundance of Faecalibacterium and Butyricicoccus were significantly increased in the G→B. With consistency in the upregulation of antioxidant enzyme ability and downregulation of pro-inflammation levels in the cecum, we observed an increase in the number of goblet cells, secretory IgA+ cells, tight junction protein (occludin, ZO-1, and claudin-1) and MUC-2 expression in the cecum of the G→B. The metabolomics analysis revealed that the relative abundance of metabolites related to butyrate was significantly increased in G→B. In an in vitro experiment, we found that butyrate could effectively induce T lymphocyte proliferation and cyclin D1 protein expression. However, these butyrate responses were abrogated by HDAC3 agonists, STAT3 antagonists, or mTOR antagonists but were mimicked by GPR43 agonists or HDAC3 antagonists. Thus, we suggested that G→B can indirectly affect the composition of cecal microbiota as well as increase the relative abundance of Faecalibacterium and Butyricicoccus and butyrate production by reducing the level of oxidative stress in the cecum. Exogenous butyrate could promote the T lymphocyte proliferation of cecal tonsil by activating the GPR43/HDAC3/p-STAT3/mTOR pathways.
Project description:Heat stress (HS) can damage the integrity of the intestinal mucosal barrier, leading to decreased poultry productivity. This study aimed to identify candidate genes related to acute HS in breeder hens and provide insight into the molecular mechanisms underlying acute HS in gut health. Fifty 28-week-old breeder hens were divided into two groups (25 hens each) raised under thermoneutral zone (23 °C) and acute HS (36 °C, 6 hours) conditions. The heart rate and cloacal temperature were measured in all hens, and jejunal mucosa tissues were randomly collected from 12 hens per treatment for RNA-sequencing analysis. The results indicated a significant increase in the heart rate and cloacal temperature in hens exposed to acute HS (P < 0.05). Transcriptome analysis identified 138 differentially expressed genes (DEGs) in heat-stressed breeder hens, including 75 upregulated DEGs containing heat shock protein (HSP), energy homeostasis metabolism-related gene (PDK4), and fat metabolism-related genes (PPARA and CD36), and 63 downregulated DEGs containing the bile acid transporter gene (SLC10A2). Gene ontology analysis revealed significant enrichment in biological processes related to heat response and cholesterol biosynthesis. Kyoto Encyclopedia of Genes and Genomes analysis highlighted several significant pathways, including steroid biosynthesis, steroid hormone biosynthesis, protein processing in endoplasmic reticulum, the peroxisome proliferator-activated receptor (PPAR) signaling pathway, and the adipocytokine signaling pathway. Protein-protein interaction network analysis involving two large networks: one containing several upregulated HSPs and genes related to energy homeostasis and fat metabolism (PDK4, PPARA, and CD36) and glucose transporter (SLC2A5), and the other containing downregulated DEGs related to cholesterol biosynthesis. Overall, acute HS might affect energy metabolism, fat metabolism, and glucose transport in the jejunal mucosa of breeder hens. Heat-stressed hens could restore the nutritional function of the jejunal mucosa by increasing the expression of HSPs. These findings provide a theoretical framework for further investigation into the molecular regulatory mechanisms responsible for HS-induced changes in the gut health of poultry.
Project description:Calcium (Ca) and phosphorus (P) are essential micronutrients linked to arrays of biological processes and physiological conditions. In laying hens, the optimal Ca/P ratio in feed is inconsistent but necessary for reliable schemes of mineral restriction in poultry diets. This study investigates the effects of dietary treatments varying in the Ca and P levels in two laying hen strains (Lohmann Brown-Classic and Lohmann LSL-Classic) at the peak of egg production (31 weeks of age). Four dietary treatment groups were differed in Ca (recommended vs. 15 % reduction) and mineral P (adequate vs. 20 % reduction) levels; 1) control diet (Con; Ca=34.4g/kg, P=5.3 g/kg and Ca/P ratio=7.45), 2) Low Ca and P diet (LCaP), 3) low Ca diet (LCa), and 4) low P diet (LP). microRNA expression of the jejunum mucosa were profiled by microRNA sequencing in a total of 80 animals (10 hens per experimental diet group for each of the two laying line) at sampling age of 31 weeks. RNA-seq data of matched samples are also available (E-MTAB-9109).
Project description:Sperm storage tubules (SSTs) in the uterovaginal junction (UVJ) of the oviduct are major sites of sperm storage after artificial insemination or mating. Female birds may regulate sperm motility in the UVJ. Heat stress can decrease the reproductive ability of broiler breeder hens. However, its effects on UVJ remain unclear. Changes in gene expression aid in understanding heat stress-affected molecular mechanisms. Herein, we wanted to conduct a comparative transcriptomic analysis to identify the differentially expressed genes (DEGs) in the UVJ of breeder hens under thermoneutral (23°C) and heat stress (36°C for 6 h) conditions.
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:The liver plays a critical role in avian reproduction as it is the primary site of de novo lipogenesis and yolk precursor synthesis. Broiler breeder hens, the parents of commercial broiler chickens, often experience poor reproductive efficiency primarily due to declining egg production beginning around 45 weeks of age. Metformin, an antidiabetic drug, exerts its primary effects in the liver by increasing insulin sensitivity and reducing hepatic glucose production in humans. This study aimed to characterize the liver transcriptomic profile of broiler breeder hens supplemented with metformin in the diet at 0 or 75 mg/kg body weight for 40 weeks (25-65 weeks of age; n=45 hens/treatment). Liver tissue was collected from a subset of hens (n=12 hens/treatment group) at 65 weeks of age, RNA was extracted and sequenced using next-generation sequencing. Differential gene abundance analysis revealed that metformin treatment led to the most significant changes in gene expression, with 552 genes differentially expressed compared to the control group. Further transcriptomic analysis highlighted increased expression of genes related to estrogen-stimulated yolk precursor synthesis, insulin-stimulated de novo lipogenesis, and AMPK-mediated glucose homeostasis. qPCR analysis revealed increased expression of ESR1, APOB, APOV1, VTG2, ADIPOQ, ADIPOR2 and ACACA mRNA and decreased expression of PCK1 mRNA validating the transcriptomic data. Collectively, the present study suggests that metformin supplementation supports prolonged egg production in aging broiler breeder hens by sustaining yolk precursor and fatty acid synthesis that are typically diminished in aging broiler breeder hens.
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.