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Next Generation Sequencing Facilitates Quantitative Analysis of Endometrium Transcriptomes During Porcine Embryo Implantation.

ABSTRACT: In this study, deep sequencing of endometrial RNA samples during early pregnancy were performed using RNA sequencing (RNA-Seq). The changes of genes expressed among three stages (day 12, 18 and 25) of pregnancy were investigated by the multiple comparisons. Analysis of reads, between 1557 and 8951 differentially expressed genes (DEGs) were revealed among different periods of implantation. Expression of selected genes was validated by the use of quantitative real-time RT-PCR. Bioinformatics analysis of differentially expressed genes in the endometrium revealed a number of biological processes and pathways potentially involved in embryo implantation in the pig, most noticeably cell proliferation, regulation of immune response, interaction of cytokine-cytokine receptor, and cell adhesion. This revealed specific gene expression patterns reflecting the different functions of the endometrium on three stages (maternal recognition, conceptus attachment and embryo implantation). This study identified comprehensive transcriptomic profile in the porcine endometrium, it could be a foundation for targeted studies of genes and pathways potentially involved in abnormal endometrial receptivity and embryo loss in early pregnancy.

ORGANISM(S): Sus scrofa

PROVIDER: GSE73695 | GEO | 2015/10/03

SECONDARY ACCESSION(S): PRJNA297678

REPOSITORIES: GEO

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Project description:Infertility and subfertility represent major problems in domestic animals and humans, and the majority of embryonic loss occurs during the first month of gestation that involves pregnancy recognition and conceptus implantation. The critical genes and physiological pathways in the endometrium that mediate pregnancy establishment and success are not well understood. In Study One, 270 predominantly Angus heifers were classified based on fertility using four rounds of serial embryo transfer (ET) to select animals with intrinsic differences in pregnancy loss. In each round, a single in vitro-produced high-quality embryo was transferred into heifers on day 7 post-estrus and pregnancy was determined on days 28 and 42 by ultrasound and then terminated. Heifers were classified based on pregnancy success as high fertile (HF), subfertile (SF), or infertile (IF). In Study Two, fertility-classified heifers were resynchronized and bred with semen from a single high fertility bull. Blood samples were collected every other day from days 0 to 36 post-mating. Pregnancy rate was determined on day 28 by ultrasound and tended to be higher in HF (70.4%) and SF (46.7%) than IF (0%) heifers. Progesterone concentrations in serum during the first 20 days post-estrus were not different in non-pregnant heifers and also not different in pregnant heifers among fertility groups. In Study Three, a single in vivo-produced embryo was transferred into fertility-classified heifers on day 7 post-estrus. The uteri were flushed on day 14 to recover embryos, and endometrial biopsies were obtained from the ipsilateral uterine horn. Embryo recovery rate and conceptus length and area were not different among the heifer groups. RNA was sequenced from the day 14 endometrial biopsies of pregnant HF, SF and IF heifers (n=5 per group) and analyzed by edgeR robust analysis. There were 26 differentially expressed genes (DEG) in the HF compared to SF endometrium, 12 DEG for SF compared to IF endometrium, and 3 DEG between the HF and IF endometrium. Many of the DEG encoded proteins involved in immune responses and are expressed in B cells. Results indicate that pre-implantation conceptus survival and growth to day 14 is not compromised in SF and IF heifers. Thus, the observed difference in capacity for pregnancy success in these fertility-classified heifers is manifest between days 14 and 28 when pregnancy recognition signaling and conceptus implantation must occur for the establishment of pregnancy. Endometrial biopsies were subjected to RNA sequencing from high fertile (HF; n=5), subfertile (SF; n=5) and infertile (IF; n=5) classified heifers on day 14 of pregnancy.

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Next Generation Sequencing Facilitates Quantitative Analysis of Endometrium Transcriptomes During Porcine Embryo Implantation.

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