Post-transfer outcomes in cultured bovine embryos supplemented with epidermal growth factor, fibroblast growth factor 2, and insulin-like growth factor 1.

This work examined the downstream fetal and placental outcomes of introducing a cocktail of uterine-derived growth factors during bovine embryo culture. Abattoir-derived bovine oocytes were matured and fertilized in vitro. On day 4 post-fertilization, ≥ 8-cell embryos were harvested, pooled and exposed to an embryokine mix, termed EFI, which contained recombinant human epidermal growth factor (10 ng/ml), bovine fibroblast growth factor-2 (10 ng/ml) and human insulin-like growth factor 1 (50 ng/ml) or to a carrier-only control treatment (CON). On day 7, individual, transfer-quality embryos were transferred to recipients. Timed ovulation was completed in mature, non-suckled commercial beef cows. Cows either were artificial inseminated (AI) or received an embryo (ET) on day 7 post-estrus (n = 23-31 cows/treatment over 4 replicate studies). The percentage of grade 1 and 2 morulae and blastocysts was greater (P < 0.05) for EFI-treated embryos than CON. The percentage of pregnant cows diagnosed by transrectal ultrasonography did not differ among the AI and ET groups on days 28, 42 and 56 post-estrus. There also were no differences in the ratio of male to female fetuses determined on day 60 post-estrus by transrectal ultrasonography. On day 21 post-estrus, the relative abundance of three interferon-stimulated gene (ISG) transcripts in peripheral leukocytes were not different based on AI/ET group or the sex of the conceptus. Circulating pregnancy-associated glycoprotein (PAG) concentrations differed (P < 0.05) among days. Also, a difference in PAG concentrations (P < 0.05) were detected between male and female pregnancies in the CON-ET group but not in the AI or EFI-ET groups. Crown-rump length was not affected by AI/ET group on day 42 but were less (P < 0.05) in the CON and EFI-ET groups than the AI group on day 56. These findings implicate EFI supplementation as a means for improving transferable embryo production in a bovine IVP system, but it is not clear if this treatment improves embryo competency after ET.

Exposure to maternal obesity alters gene expression in the preimplantation ovine conceptus.

BACKGROUND: Embryonic and fetal exposure to maternal obesity causes several maladaptive morphological and epigenetic changes in exposed offspring. The timing of these events is unclear, but changes can be observed even after a short exposure to maternal obesity around the time of conception. The hypothesis of this work is that maternal obesity influences the ovine preimplantation conceptus early in pregnancy, and this exposure will affect gene expression in embryonic and extraembryonic tissues. RESULTS: Obese and lean ewe groups were established by overfeeding or normal feeding, respectively. Ewes were then bred to genetically similar rams. Conceptuses were collected at day 14 of gestation. Morphological assessments were made, conceptuses were sexed by genomic PCR analysis, and samples underwent RNA-sequencing analysis. While no obvious morphological differences existed between conceptuses, differentially expressed genes (≥ 2-fold; ≥ 0.2 RPKM; ≤ 0.05 FDR) were detected based on maternal obesity exposure (n = 21). Also, differential effects of maternal obesity were noted on each conceptus sex (n = 347). A large portion of differentially expressed genes were associated with embryogenesis and placental development. CONCLUSIONS: Findings reveal that the preimplantation ovine conceptus genome responds to maternal obesity in a sex-dependent manner. The sexual dimorphism in response to the maternal environment coupled with changes in placental gene expression may explain aberrations in phenotype observed in offspring derived from obese females.

Maternal obesity alters the expression of embryonic regulatory transcripts in the preimplantation ovine conceptus.

The influence of exposure to overfeeding-induced maternal obesity around the time of conception on early embryogenesis was examined in the day 14 ovine conceptus. The relative abundance of FGFR2 and DNMT1 was influenced by maternal obesity status and conceptus sex, and the abundance of PPARG and PTGS2 transcripts was greater in male conceptuses regardless of the obesity status of the ewe. These observations demonstrated that short-term exposure to maternal obesity impacts early conceptus transcript patterning.