Expression data from early luteal bovine endometrium
Ontology highlight
ABSTRACT:
ORGANISM(S): Bos taurus
SUBMITTER: aideen killeen
PROVIDER: E-GEOD-29853 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:The aim of this study was to examine the effect of sire fertility status on conceptus-induced changes in the endometrial transcriptome. Holstein Friesian bulls (3 High fertility, HF, 3 Low fertility, LF) were selected from the Irish national population of AI bulls (minimum of 500 inseminations/bull) based on adjusted fertility scores (HF: +4.37% and LF: -12.7%; mean = 0%). To generate elongated conceptuses, Day 7 blastocysts produced in vitro using sperm from these six bulls were transferred in groups of 5-10 to synchronized heifers (n=7 heifers per bull; total 42 heifers). Conceptuses were recovered following slaughter on Day 15 (recovery rate: HF 59.4% vs. LF 45.0%; P<0.05). In parallel, Day 15 endometrial explants were recovered from synchronized cyclic heifers (n=4). Explants from each heifer were co-cultured for 6 h in RPMI medium with (i) nothing, control (ii) 100 ng/ml ovine recombinant interferon tau (IFNT) (iii) a single conceptus from each high fertility bull, or (iv) a single conceptus from each low fertility bull. To minimize variation, explants from the same uterus were used across all treatments, replicated across 4 heifers. After 6 h, explants were snap frozen and stored at -80°C. Extracted mRNA was subjected to RNA-seq (Illumina NextSeq 500) and the resulting data were analyzed through a bioinformatic pipeline with R software.
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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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Project description:The aberrant gene expression in the uterine endometrium and embryo has been the major causes of pregnancy failure in cattle. Therefore, selecting cows having adequate endometrial receptivity and embryos of better developmental competence based on the gene expression could increase the number of calves produced by each cow during its productive life time. We used endometrial and embryo biopsy technology in conjunction with the pregnancy outcome information to establish a direct link between the pre-transfer endometrial or in vivo derived embryo gene expression and pregnancy outcome after embryo transfer. Endometrial samples were collected from Simmental heifers at day 7 and 14 of the estrous cycle, one cycle prior to embryo transfer. In the next cycle, embryo biopsies consisting of 60-70% of inner cell mass and trophectoderm were transferred to the recipients at day 7 of the estrous cycle. The remaining 30-40% parts of the embryos were retained for analysis.Pregnancy diagnosis was performed at days 28 and 42 by ultrasonography and at day 56 by rectal palpation. Those heifers returned to heat at day 21 were considered as non pregnant or non receptive endometrium (NP) while those heifers ended up with successful pregnancy and calf delivery was considered as the calf delivery group or receptive endometrium (CD). Following this, the endometrial samples collected during the pre-transfer period and the embryo biopsies retained during embryo transfer were categorized based on the pregnancy outcome. Those endometrial biopsies collected at days 7 and 14 of the estrous cycle from heifers resulted in successful calf delivery were designated as CDd7 and CDd14, respectively and endometrial biopsies taken at days 7 and 14 of the estrous cycle from those subsequently resulted in no pregnancy were designated as NPd7 and NPd14, respectively. Similarly, the embryo biopsies were classified as those embryo biopsies resulted in successful calf delivery and those resulted in no pregnancy
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Project description:To identify heifers of contrasting fertility, serial rounds of artificial insemination (AI) were conducted in 201 synchronized crossbred beef heifers. The heifers were then fertility classified based on number of pregnancies detected on day 35 in the four AI opportunities. The reproductive tracts of a subset of the classified heifers were obtained on day 14 of the estrous cycle. Microarray analysis revealed differences in the endometrial transcriptome based on fertility classification.
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