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 AlexaM-BM-. 555 fluorescent dye and the other half with AlexaM-BM-. 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: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
Project description:cDNA microarrays were used to identify the expression of the genes in uterus by comparison with two others regions of the oviduct not involved in the shell deposition. A total of 605 sequences specifically over-expressed were revealed. The expression of 15 of them was also confirmed using qRT-PCR, and confirmed the microarray expression. The 605 uterine transcripts corresponded to 469 and 437 different genes and proteins respectively. A global interpretation of their functions was assessed using the Gene Ontology terms and the most over-represented terms are relative to the mineral transport and transfer necessary to the calcification process. The study has revealed 53 proteins which could be secreted to be deposited in the shell to be biologically actives. The study of their functions reveals proteins known for their involvement in the biomineralization or with calcium binding properties to interact with the mineral phase during the eggshell fabric. A large group of proteins could be involved in the folding of the matrix, and a notable group of uterine secreted proteins was also identified as antibacterial to contribute to the antimicrobial protection of the egg. Finally the study revealed proteases and protease inhibitors which could play an active role in the regulation of the protein activities in the acellular uterine fluid where the eggshell is formed. Keywords: Laying hen, eggshell, oviduct, uterine expression, cDNA microarray, indirect cDNA labelling, Alexa Fluor dyes 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 (uterus versus magnum and uterus versus white isthmus).
Project description:Expression of known and predicted genes in tissues of Gallus gallus (chicken) pooled from multiple healthy individuals. Two-colour experiments with two different tissues hybridized to each array. Each tissue is arrayed in replicate with dye swaps. Tissues: Bursa of Fabricius, Cerebellum, Cerebral cortex, Eye, Femur with bone marrow, Gallbladder, Gizzard, Heart, Intestine, Kidney, Liver, Lung, Muscle, Ovary, Oviduct, Skin, Spleen, Stomach, Testis, Thymus
Project description:Hypoxia is a condition in which tissues of the body do not receive sufficient amounts of oxygen supply. The current study aimed to clarify the impact of a forced low-oxygen environment on the early pregnancy by exposing mice to low-oxygen conditions for 72 hours after fertilization. The treatment resulted in a complete failure of blastocyst implantation accompanied by vascular hypermeability in the uterus. The transcriptome analysis of uterus and ovary revealed remarkable alterations in gene expression between the control normoxic and hypoxic treatment groups. These alterations were encompassed a number of genes categorized to the immune responses and oxidative demethylation process.