Publications by authors named "Alan D Ealy"

Article Synopsis
  • This study investigated the role of interleukin-6 (IL6) cytokine signaling in bovine embryo development by inhibiting the IL6 signal transducer (IL6ST).
  • Using a pharmacological inhibitor (SC144), the researchers found that blocking IL6ST signaling diminished embryo development and reduced cell numbers in critical stages like the 16-cell and blastocyst stages.
  • Additionally, employing CRISPR-Cas9 to disrupt IL6ST showed high editing efficiency and similarly affected embryo development, indicating that IL6 family signals are crucial for normal bovine embryo growth and organization during these early stages.
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This work explored whether a well-characterized recombinant human interleukin-6 (hIL6) protein will influence in vitro produced (IVP) bovine embryo development and survival after cryopreservation. Cumulus oocyte complexes were collected from abattoir derived ovaries, matured for 24 h, and fertilized using pooled semen from Holstein bulls. Embryos were treated with 0, 25, 50, or 100 ng/mL hIL6 on day 5 post-fertilization.

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A high incidence of pregnancy failures occurs in cattle during the second week of pregnancy as blastocysts transition into an elongated conceptus. This work explored whether interleukin-6 supplementation during in vitro embryo production would improve subsequent conceptus development. Bovine embryos were treated with 0 or 100 ng/mL recombinant bovine interleukin-6 beginning on day 5 post-fertilization.

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Article Synopsis
  • In vitro production (IVP) of bovine embryos is increasingly preferred over conventional methods due to its effectiveness with genetically elite heifers and cows.
  • However, low efficiency and challenges like the impact of reactive oxygen species (ROS) negatively affect embryo quality during the IVP process.
  • This review will discuss how adding antioxidant supplements can improve various stages of embryo development, including oocyte maturation, sperm viability, fertilization, and culture.
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This work explored whether supplementing selective members of the interleukin-6 (IL6) cytokine family during in vitro bovine oocyte maturation affects maturation success, cumulus-oocyte complex (COC) gene expression, fertilization success, and embryo development potential. Human recombinant proteins for IL6, IL11, and leukemia inhibitory factor (LIF) were supplemented to COCs during the maturation period, then fertilization and embryo culture commenced without further cytokine supplementation. The first study determined that none of these cytokines influenced the rate that oocytes achieved arrest at meiosis II.

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CRISPR-Cas ribonucleoproteins (RNPs) are important tools for gene editing in preimplantation embryos. However, the inefficient production of biallelic deletions in cattle zygotes has hindered mechanistic studies of gene function. In addition, the presence of maternal RNAs that support embryo development until embryonic genome activation may cause confounding phenotypes.

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Article Synopsis
  • Early embryonic pregnancy losses in beef and dairy cows are often linked to developmental issues occurring during various stages, especially with in-vitro-produced embryos.
  • The review covers critical stages like oocyte maturation, fertilization, and early embryonic development, highlighting common problems encountered and their impact on pregnancy outcomes.
  • While some potential solutions are explored, many issues currently lack effective corrective measures, indicating a need for further research to enhance the survival rates of bovine embryos to term.
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Early embryonic mortality caused by maternal-fetal recognition failure in the three weeks after fertilization represents a major cause of reproductive inefficiency in the cattle industry. Modifying the amounts and ratios of prostaglandin (PG) F and PGE can benefit the establishment of pregnancy in cattle. Adding conjugated linoleic acid (CLA) to endometrial and fetal cells culture affects PG synthesis, but its effect on bovine trophoblast cells (CT-1) is unknown.

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Cattle embryos represent a useful model for understanding parts of human embryogenesis due to various biological similarities. We describe a protocol to mature and fertilize bovine oocytes followed by culture of resulting presumptive zygotes up until the blastocyst stage. Our protocol features a unique procedure for washing and moving oocytes and zygotes between their respective dishes using a cell strainer.

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Article Synopsis
  • In vitro production of embryos (IVP) aims to create high-genetic-value embryos, but current culture systems have low efficiency levels.
  • This study explored two selection methods—BCB staining and cleavage kinetics—to better identify embryos with higher developmental potential.
  • Results showed that while BCB staining improved blastocyst development, using both BCB and cleavage kinetics together negated the benefits of BCB alone; cleavage kinetics alone resulted in the best rate of blastocyst development but didn't predict embryo survival after cryopreservation.
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Background: Cytoplasmic and nuclear maturation of oocytes, as well as interaction with the surrounding cumulus cells, are important features relevant to the acquisition of developmental competence.

Methods: Here, we utilized Brilliant cresyl blue (BCB) to distinguish cattle oocytes with low activity of the enzyme Glucose-6-Phosphate Dehydrogenase, and thus separated fully grown (BCB positive) oocytes from those in the growing phase (BCB negative). We then analyzed the developmental potential of these oocytes, mitochondrial DNA (mtDNA) copy number in single oocytes, and investigated the transcriptome of single oocytes and their surrounding cumulus cells of BCB positive versus BCB negative oocytes.

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Ovum pickup and in vitro production (IVP) of bovine embryos are replacing traditional multiple ovulation embryo transfer (MOET) as the primary means for generating transferable embryos from genetically elite sires and dams. However, inefficiencies in the IVP process limit the opportunities to produce large numbers of transferable embryos. Also, the post-transfer competency of IVP embryos is inferior to embryos produced by artificial insemination or MOET.

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Article Synopsis
  • Fetal loss in cattle is linked to issues during oocyte maturation, embryonic development, and communication between the conceptus (early embryo) and the endometrium (lining of the uterus), leading to pregnancy failure.
  • The review explores how paracrine signaling, especially involving interferon-tau and other secretory factors from both the conceptus and endometrium, influences pregnancy establishment and maintenance.
  • Recent advancements in "omic" technologies reveal that the endometrium can respond differently based on conceptus sex and origin, highlighting the need for further research to reduce early embryonic loss in cattle and enhance reproductive success.
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Exposure to maternal obesity in utero is associated with marked developmental effects in offspring that may not be evident until adulthood. Mechanisms regulating the programming effects of maternal obesity on fetal development have been reported, but little is known about how maternal obesity affects the earliest periods of embryonic development. This work explored how obesity influences endometrial gene expression during the peri-implantation period using a sheep model.

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The term "embryokine" has been used to denote molecules produced by the endometrium, oviduct, or by embryo itself that will influence embryo development. Several cytokines have been identified as embryokines in cattle and other mammals. This review will describe how these cytokines function as embryokines, with special emphasis being placed on their actions on in vitro produced (IVP) bovine embryos.

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Body systems once thought sterile at birth instead have complex and sometimes abundant microbial ecosystems. However, relationships between dam and calf microbial ecosystems are still unclear. The objectives of this study were to (1) characterize the various maternal and calf microbiomes during peri-partum and post-partum periods and (2) examine the influence of the maternal microbiome on calf fecal microbiome composition during the pre-weaning phase.

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The objective of the study was to examine how l-citrulline supplementation to ewes during mid-gestation influences placental activity, placental blood flow, lamb body weight, and carcass characteristics. Two studies were completed. A pharmacokinetic study to compare circulating plasma amino acid concentrations after a single intravenous injection of 155 µmol/kg BW l-citrulline or after an isonitrogenous amount of l-alanine (control; 465 µmol/kg BW).

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The use of in vitro produced embryos in dairy and beef cattle has increased in recent years, but compromised post-transfer pregnancy success prevents producers from capturing all the benefits this technology can provide. This study explored whether supplementing interleukin-6 (IL6) during in vitro embryo development influences post-transfer development of the embryo-proper, fetus and placenta during early gestation in cattle. Slaughterhouse-derived cumulus oocyte complexes underwent IVM (day -1) and IVF (day 0).

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The worldwide production of in vitro-produced embryos in livestock species continues to grow. The current gold standard for selecting quality oocytes and embryos is morphologic assessment, yet this method is subjective and varies based on experience. There is a need for a non-invasive, objective method of selecting viable oocytes and embryos.

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Background: Interleukin-6 (IL6) was recently identified as an embryotrophic factor in bovine embryos, where it acts primarily to mediate inner cell mass (ICM) size. This work explored whether IL6 affects epiblast (EPI) and primitive endoderm (PE) development, the two embryonic lineages generated from the ICM after its formation. Nuclear markers for EPI (NANOG) and PE (GATA6) were used to differentiate the two cell types.

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Deficiencies in current embryo culture media likely contribute to the poor blastocyst development rates and pregnancy retention rates for in vitro produced (IVP) bovine embryos. Of special concern is the lack of micronutrients in these media formulations. One micronutrient of interest is zinc, an essential trace element involved with various enzyme and transcription factor activities.

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Several tools exist to diagnose pregnancy in dairy cattle. However, substantial pregnancy loss occurs within the first 60 d of gestation in cattle, and these losses have a profound adverse economic impact on the dairy and beef cattle industries. Detecting these impending pregnancy losses could offer producers an opportunity to reduce costs associated with this source of reproductive inefficiency.

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Article Synopsis
  • Supplementing interleukin-6 (IL6) in cultured bovine embryos boosts the number of inner cell mass (ICM) cells in blastocysts, while showing no effect on morulae cell numbers.
  • IL6 triggers phosphorylation and nuclear movement of a protein called STAT3 in early morulae and ICM cells of blastocysts, and it also increases the gene SOCS3, which is responsive to STAT3.
  • Blocking Janus kinase (JAK) activity prevents IL6 from activating STAT3, leading to a decrease in ICM cell numbers, highlighting IL6’s role as a key regulator during early blastocyst development.
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