Whole genome bisulfite sequencing of bovine oocytes from metabolically divergent lactating cows and nulliparous heifers
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PROVIDER: PRJEB33720 | ENA |
REPOSITORIES: ENA
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Project description:Lactating cows experience transient metabolic stresses during postpartum which results in abnormal concentrations of non-esterified fatty acids and beta-hydroxybutyrate that can impact oocyte maturation and ultimately reproductive success. We hypothesize that metabolic challenges in lactating cows influence DNA methylation changes of the oocyte affecting genes involved in oocyte developmental competency. Therefore, we compared whole genome bisultfite sequencing (wgbs) of the oocytes collected from 5-6 week (early) or 9-10 week (mid) post-partum against that of oocytes from nulliparous heifers as metabolically unchallenged condition control. Bisulfite-Seq DNA libraries were generated from pooled oocytes using an EZ DNA methylation-direct kit and Pico-Methyl seq library preparation kit (Zymo Research), and parallel sequenced using the illumina HiSeq2500 system.
2021-11-30 | E-MTAB-8191 | biostudies-arrayexpress
Project description:Lactating cows experience transient metabolic stresses during postpartum which results in abnormal concentrations of non-esterified fatty acids and beta-hydroxybutyrate that can affect embryo-maternal communication and ultimately reproductive success. We hypothesize that metabolic challenges in lactating cows influence DNA methylation changes of the embryos prior to implantation affecting genes involved in embryo developmental competency. Therefore, we compared whole genome bisulfite sequencing (wgbs) of morulas derived from in vitro produced 2-4 cell embryos and transferred in the oviduct of metabolically profiled lactating cows until day 7. Similar stage of in vitro production (IVP) embryos were transferred to the oviduct of nulliparous heifers as a metabolically unchallenged control. Bisulfite-Seq DNA libraries were generated from groups of five morulas using an EZ DNA methylation-direct kit and Pico-Methyl seq library preparation kit (Zymo Research), and parallel sequenced using the illumine HiSeq2500 system at the institute of Genome Biology, Leibniz-Institute for Farm Animal Biology (FBN), Dummerstorf, Germany.
2021-11-30 | E-MTAB-8239 | biostudies-arrayexpress
Project description:Infertility in lactating dairy cows is explained partially by the metabolic state associated with high milk production. The hypothesis was that lactating and non-lactating cows would differ in endometrial and placental transcriptomes during early pregnancy (day 28 to 42) and this difference would explain the predisposition for lactating cows to have embryonic loss at that time. Cows were either milked or not milked after calving. Reproductive [endometrium (caruncular and intercarunclar) and placenta] and liver tissues were collected on day 28, 35, and 42 of pregnancy. The primary hypothesis was rejected because no effect of lactation on mRNA abundance within reproductive tissues was found. Large differences within liver demonstrated the utility of the model to test an effect of lactation on tissue gene expression. Major changes in gene expression in reproductive tissues across time were found. Greater activation of the transcriptome for the recruitment and activation of macrophages was found in the endometrium and placenta. Changes in glucose metabolism between day 28 and 42 included greater mRNA abundance of rate-limiting genes for gluconeogenesis in intercaruncular endometrium and evidence for the establishment of aerobic glycolysis (Warburg effect) in the placenta. Temporal changes were predicted to be controlled by CSF1, PDGFB, and JUN. Production of nitric oxide and reactive oxygen species by macrophages was a mechanism to promote angiogenesis in the endometrium. Reported differences in pregnancy development for lactating versus non-lactating cows could be explained by systemic glucose availability to the conceptus and appear to be independent of the endometrial and placental transcriptomes.
2017-08-10 | GSE94794 | GEO
Project description:The aim of this study was to evaluate the potential effects of methyl donor supplementation of pregnant animals in the presence or absence of a concomitant lactation on the methylome of the offspring. Twenty Holstein cows, 10 nulliparous (not lactating while pregnant) and 10 multiparous (lactating while pregnant) were blocked by parity and randomly assigned to an i.m. weekly injections of a placebo (CTRL) or a solution containing methyl donors (MET). After calving, 5 calves randomly selected from each treatment (two born to primiparous and three to multiparous dams) were blood-sampled to determine their full methylome. There were more than 2,000 CpG differentially methylated between calves born to CTRL and those born to MET, and also between calves born to multiparous and nulliparous dams. Most of the differences affected genes involved in immune function, cell growth regulation and differentiation, kinase activity, and ion channeling. We conclude that the coexistence of pregnancy and lactation affects the methylome of the offspring, and that supplementation of methyl donors early in gestation has also consequences on the methylome.
2017-08-17 | GSE102736 | GEO
Project description:Whole genome bisulfite sequencing of bovine morula derived from metabolically divergent dairy cows
| PRJEB33956 | ENA
Project description:Faecal samples from lactating dairy cows Raw sequence reads
| PRJNA449795 | ENA
Project description:The hypothesis tested was that the uterine environment of lactating cows would affect conceptus gene expression. Approximately 65-75 days post-partum (dpp) the estrous cycles of non-lactating (dried off immediately post partum: n=12) and lactating (n=13) cows were synchronized and on Day 7 a high quality blastocyst derived from superovulated heifers was transferred. A control group of maiden heifers (n=8) were synchronized, inseminated to a standing heat and slaughtered on the same day as non-lactating and lactating recipients (Day 19; estrus=Day 0). The ipsilateral uterine horn was flushed with 10 ml PBS and the conceptus, when present, and uterine luminal fluid (ULF) snap frozen in liquid nitrogen prior to analysis. Gene expression analysis of the conceptus was performed by RNA sequencing analysis while amino acid (aa) composition of ULF was determined by High Performance Liquid Chromatography (HPLC). Eight differentially expressed genes (DEGs) were identified between conceptuses recovered from non-lactating cows versus heifers while 269 DEGs (100 up-regulated and 169 down-regulated) were identified between conceptuses recovered from lactating cows compared to heifers. The aa, alanine, glycine, serine and threonine, arginine, leucine and valine, were significantly lower in abundance in ULF recovered from heifers compared to both non-lactating or lactating cows. Glutamic acid, glutamine and lysine concentrations were lowest in heifers compared to both cow groups. This study demonstrates that exposure of a grade one embryo to a uterine environment that has been exposed to the metabolic stresses associated with lactation modifies the transcriptome of the conceptus and aa composition of the ULF.
2017-08-12 | GSE85563 | GEO
Project description:Evaluate the influence of maternal metabolism on gene expression profiles from extra-embryonic tissues at D18 Keyword: Holstein Heifers and Postpartum dairy cows, metabolism and energy status, elongation and gastrulation, extra-embryonic tissues, transcriptome, correlations between genes & metabolites?
2014-02-04 | GSE46940 | GEO
Project description:Evaluate the influence of maternal metabolism on gene expression profiles from extra-embryonic tissues at D18 Keyword: Holstein Heifers and Postpartum dairy cows, metabolism and energy status, elongation and gastrulation, extra-embryonic tissues, transcriptome, correlations between genes & metabolites? 8 replicates in GH, 11 replicates in ELC and 10 replicates in LLC (extra-embryonic tissues) per group of female (N=3) were hybridized (total N=29) against a placental reference that hybridized itself to 99% of the array. Hybridization design: 6 microarrays per experiment (N=5) were randomly hybridized to the EET from 2 conceptuses originating either from development in heifers, ELC or LLC. Extra-embryonic tissues were systematically labelled with Cy3 and Reference in Cy5.
2014-02-04 | E-GEOD-46940 | biostudies-arrayexpress
Project description:Cellular mechanisms that contribute to low estradiol concentrations produced by the preovulatory ovarian follicle in cattle with a compromised metabolic status (such as lactatino) are largely unknown. To gain insight into the main metabolic mechanisms affecting preovulatory follicle function RNAseq profiling was conducted on non-lactating Holstein-Friesian heifers (n=16) and lactating Holstein-Friesian cows (n=17) at three stages of preovulatory follicle development: A) newly selected dominant follicle in the luteal phase (Selection); B) follicular phase before the LH surge (Differentiation) and C) pre-ovulatory phase after the LH surge (Luteinization). Based on a combination of RNA sequencing, ingenuity pathway analysis and Q-RT-PCR validation several important molecular markers involved in steroid biosynthesis, such as the expression of steroidogenic acute regulatory protein (STAR) within developing dominant follicles, were identified to be affected (downregulated) by the catabolic state. We propose that the adverse metabolic environment caused by lactation decreases preovulatory follicle function by affecting cholesterol transport into the mitochondria to initiate steroidogenesis. Granulosa and Theca samples from the dominant follicle were taken from cows and heifers at stages: selection, differentiation and luteinization.
2012-06-15 | E-GEOD-34317 | biostudies-arrayexpress