Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of pig endometrium from pregnant and non-pregnant sows

ABSTRACT: In an attempt to unveil part of the molecular processes controlling porcine placentation we have investigated the pregnancy induced gene expression in the porcine endometrium at Day 14 after insemination using the Affymetrix GeneChip® Porcine Genome Array. Experiment Overall Design: At Day 14 after insemination, samples were obtained from the endometrium of pregnant sows and sows inseminated with inactivated semen for RNA extraction and hybridization on Affymetrix microarrays. The uteri were removed and each uterine horn was flushed with PBS containing 1% fetal calf serum, and subsequently opened longitudinally at the anti-mesometrial side. The sites of embryonic attachment were macroscopically visual in the endometrium on the mesometrial side in form of hyperemic zones. Samples of the endometrium (lamina epithelialis, lamina propria and tela submucosa, but not tunica muscularis) were isolated from proximal (the end close to the ovaries), intermedial, and distal (next to the corpus uteri)) sections of each uterine horn. Samples were taken from the endometrium of the non-pregnant animals at comparable locations.

ORGANISM(S): Sus scrofa

SUBMITTER: Esben Oestrup

PROVIDER: E-GEOD-18641 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The porcine conceptus undergoes rapid differentiation and expansion of its trophoblastic membranes between Days 11 and 12 of gestation. Concomitant with trophoblast elongation, production of conceptus estrogen, the porcine embryonic pregnancy recognition signal increases. Conceptus attachment to the uterine surface epithelium starts after Day 13 initiating epitheliochorial placentation. To analyze the transcriptome changes in the endometrium in the course of maternal recognition of pregnancy (MRP), deep sequencing of endometrial RNA samples of Day 12 pregnant animals (n=4) and corresponding non-pregnant controls (n=4) was performed using Illumina RNA-Seq. Between 30 and 35 million sequence reads per sample were produced and mapped to the porcine genome (Sscrofa10.2). Analysis of read counts revealed 2,593 differentially expressed genes (DEG). Expression of selected genes was validated by the use of quantitative real-time RT-PCR. Bioinformatics analysis identified several functional terms specifically overrepresented for up-regulated or down-regulated genes. Comparison of the RNA-Seq data from Days 12 and 14 of pregnancy was performed at the level of all expressed genes, of the DEG, and at the level of functional categories. This revealed specific gene expression patterns, reflecting the different functions of the endometrium during these stages, i.e. recognition of pregnancy and preparation for conceptus attachment. Genes related to mitosis, immune response, epithelial cell differentiation and development, proteolysis, and prostaglandin signaling and metabolism are discussed in detail. In conclusion, this study identified comprehensive transcriptome changes in porcine endometrium associated with MRP and initiation of implantation and a number of genes and pathways potentially involved in regulation of these processes. In total, 8 samples were analyzed, 4 biological replicates for pregnant animals (samples from 4 different animals) and 4 biological replicates for cyclic controls (samples from 4 different animals)

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Transcription profiling of pig endometrium from pregnant and non-pregnant sows

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