Project description:We used Affymetrix Chicken Genome Array to identify transcripts differentially expressed between cells treated with DMIOA and control cells and between DMIOA and DMIOA+20S treated cells Preadipocytes isolated from laying hen were treated with DMIOA and DMIOA+20S to identify transcripts differentailly expressed between control cells and DMIOA treated and between DMIOA and DMIOA+20S treated cells
Project description:The Del-Mar 14K chip was used to interrogate differential expression of transcripts in the white isthmus (WI) compared with the adjacent magnum (Mg) and uterine (Ut) segments of the hen oviduct. Differential expression of genes common to both comparisons (WI/Mg and WI/Ut) was detected for 204 annotated proteins. Of these, 58 genes were overexpressed in both WI/Mg and WI/Ut, and are therefore considered to be the most interesting candidates for WI - specific functions. Additionally, general analysis revealed 135 clones hybridizing to overexpressed transcripts (WI/Mg + WI/Ut), and corresponding to 102 NCBI annotatated non-redundant Gallus gallus gene ID~s. This combined analysis revealed that structural proteins highly over-expressed in white isthmus were collagen X (COL10A1), Fibrillin (FBN1) and Cysteine Rich Eggshell Membrane Protein (CREMP). In addition, genes encoding collagen-processing enzymes were over-expressed, as were proteins known to regulate disulfide cross-linking, suggesting that coordinated upregulation of gene networks in the white isthmus is associated with eggshell membrane fibre formation. IPA interactome analysis reinforces the key role of the estrogen receptor and SMAD3 in mediating gene regulation during eggshell membrane synthesis. These results will assist with development of selection strategies to improve eggshell quality and food safety of the table egg. Keywords: Laying hen, eggshell, oviduct, Isthmus expression, cDNA microarray, indirect cDNA labelling, Alexa Fluor dyes Keywords: Expression profiling by array A balanced block hybridization design (Dye switch) was used where half of the samples were labelled with Alexa® 555 fluorescent dye and the other half with Alexa® 647. A total of 16 microarray slides were used for hybridization to 32 samples that correspond to four tissue contrast (White isthmus versus magnum and uterus versus white isthmus).
Project description:Adipogenesis is one of the most intensively studied models of cellular differentiation, and transcriptional activities involved in the process have been extensively investigated. We analyzed transcripts differentially expressed between hen preadipocytes treated with adipogenic cocktail containing 500 nM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine, 20 µg/mL insulin and 300 μM OA (DMIOA) and non-treated control cells and transcripts differentially expressed between preadipocytes treated with DMIOA alone and those treated with a combination of DMIOA and 20(S)-hydroxycholesterol (DMIOA + 20(S)) using Affymetrix GeneChip® Chicken Genome Array containing 28,000 transcripts. Preadipocytes were isolated from hen abdominal tissue, cultured and treated with DMIOA, DMIOA+Retinoic Acid
Project description:A total of 565 miRNAs annotated in miRBase 20.0 were identified to be expressed in the liver of hen by high-throughput sequencing three biological reduplication libraries in juvenile and egg-laying hens, respectively. Compared with juvenile hen, 80 miRNAs (67 down-regulated and 13 up-regulated) were verified to be significant differential expression (SDE) in egg-laying physiological stage. Among these, miR-22-3p has the highest abundant expression, and miR-146b-5p has the highest fold-change. Additionally, 19 of the 71 novel miRNAs was significantly expressed. Furthermore, 648 putative target genes of the SDE miRNAs were obtained, and among these, FADS1, FADS2, ELOVL6, ACSL5, etc which are lipid metabolism related critical regulators are targeted by some SDE miRNAs. Gene Ontology (GO) analyses to the putative target genes of all the SDE miRNAs showed significantly enriched in Steroid biosynthesis, Glycerophospholipid metabolism, Biosynthesis of unsaturated fatty acids, and PPAR signaling pathway (P ≤ 0.05). Meanwhile, GO terms are also significantly enriched in lipid related biological processes. Overall design: Total two groups juvenile and egg-laying hens were included,and three replications for each group.Juvenile stage was designed as the reference group.The liver miRNAs expression profile of juvenile hens and laying hens were generated by Solexa microRNA-Seq
Project description:The aim of this study was to assess the impact of oocyte competence on subsequent fertility. Based on knowledge already accessible in mammals and on bioinformatics tools including the chicken genome sequence, we focused on the expression of genes involved in the processes of fertilization and of early embryo development. A differential kinetic study is performed on INRA lines selected on the basis of their fertility potential in purpose of hopefully access gene markers of fertility performance. We use 4 different hen lines: - one line of laying hens with 3 different samples: the just ovulated oocyte, the oocyte collected 24 hours before ovulation (F1 stage), and granulosa cells collected at the F1 stage. We could compare different tissue and developmental stages. - one line of hen with rapid growth speed - two lines of laying hens For the 3 last lines we used animals with different fertility levels. We collected the oocyte of the largest follicle before ovulation (F1). The aim of the study is to identify genes involved in fertility or early embryo mortality. Keywords: normal vs disease comparison Overall design: 6 arrays - Gallus gallus
Project description:Ovarian cancer has a high mortality rate due, in part, to the lack of early detection and incomplete understanding of the origin of the disease. The hen is the only spontaneous model of ovarian cancer, and can therefore aid in the identification and testing of early detection strategies and therapeutics. To our knowledge, no studies to date have examined global gene expression in ovarian cancer of the hen. Our aim was to combine the use of the hen animal model and microarray technology to identify differentially expressed genes in ovarian tissue from normal hens compared to hens with ovarian cancer. Overall design: Ovarian tissue samples from whole ovaries were collected from hens for RNA extraction and hybridization on Affymetrix microarrays. Hens were matched for age and laying status. Normal hens (n=3) showed no gross or histopathological evidence of ovarian cancer, while cancer specimens (n=3) had tumors that were stage 2 (restricted to the ovary and observable at necropsy) or 3 (ovarian tumor with abdominal seeding). Total RNA was extracted using TRIZOL according to the manufacturer's instructions.