Unknown,Transcriptomics,Genomics,Proteomics

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Whole genome bisulfite sequencing of bovine morula derived from metabolically divergent dairy cows

ABSTRACT: 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.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Bos taurus

SUBMITTER: Nares Trakooljul

PROVIDER: E-MTAB-8239 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Objective: To study post-hatching development after transfer of morphologically good quality embryos that survived in vitro maturation under lipolytic conditions using a well-established and recognized bovine model. Design: prospective study with a crossover design. Setting: research laboratory. Animals: 8 healthy multiparous non-lactating Holstein cows used in repeated embryo transfers. Intervention(s): Bovine cumulus oocyte complexes (COCs) were in vitro matured in the presence of either 1) a combination of elevated pathophysiological concentration of palmitic acid (PA) and physiological concentrations of stearic acid (SA) and oleic acid (OA) (HIGH PA), or 2) physiological concentrations of PA, SA and OA (BASAL). Matured oocytes were fertilized and cultured in FA-free conditions until day 7.7 p.i. Equal proportions of normal and expanded blastocysts were subsequently transferred to synchronized cows. Conceptuses were transcervically recovered 7 days after the transfer. Main Outcome Measure(s): Recovered embryos were morphologically assessed and dissected into embryonic disc (ED) and extraembryonic tissue (EXT). Samples of EXT were used for analysis of interferon-τ (IFN-τ) secretion and measurement of cellular metabolic activity. ED and EXT samples were subjected to RNA sequencing in order to evaluate the genome wide transcriptome patterns of recovered day 14 embryos. Results: Recovered embryos resulting from metabolically compromised oocytes were retarded in growth or less elongated than their control counterparts. Furthermore, they exhibited a decrease in glucose consumption and lactate production. Importantly, lipolytic conditions resulted in reduced capacity of the embryo to signal its presence to the mother through IFN-τ secretion. The transcriptome profile was altered in the recovered embryos with main effects in pathways related to redox-regulating mechanisms, apoptosis, cellular growth, interaction and differentiation, energy metabolism, and epigenetic mechanisms.

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Whole genome bisulfite sequencing of bovine morula derived from metabolically divergent dairy cows

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