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End of project report Embryo loss is a major cause of reproductive wastage in the cow. The majority of embryo loss occurs in the first 16 days after fertilisation when the embryo is critically dependent on the maternal uterine environment for survival. Despite the central role of uterine fluid in the normal growth and development of the embryo, there is limited information on the protein composition of these fluids. The main objectives of the studies in this thesis were to examine the protein composition of the bovine uterus during the oestrous cycle and to examine the relationship between the concentration of systemic progesterone and uterine protein expression in the cow. In the first study, the concentration of retinol-binding protein (RBP) in the bovine uterus was found to vary across the cycle and was 5-15-fold higher (P0.05) of the cycle, which was surprising given that previous studies have indicated that uterine RBP gene expression was positively associated with the concentration of systemic progesterone. In the second study, IGF binding protein 2 (IGFBP-2), IGFBP-3, IGFBP-4 and IGFBP-5 were identified in uterine fluid on days 3, 7, 11 and 15 of the oestrous cycle. There was a local effect on the concentration of IGFBPs where the concentration was greater on the ipsilateral side than that on the contralateral side for IGFBP-2 (P

End of project report Negative energy balance (NEB) is a severe metabolic affecting high yielding dairy cows early post partum with both concurrent and latent negative effects on cow fertility as well as on milk production and cow health. The seasonal nature of Irish dairy production necessitates high cow fertility and a compact spring calving pattern in order to maximise grass utilisation. Poor dairy cow reproductive performance currently costs the Irish cattle industry in excess of €400 million annually. High milk yields have been associated with lower reproductive efficiency, and it has been suggested that this effect is probably mediated through its effects on the energy balance of the cow during lactation. The modern high genetic merit dairy cow prioritises nutrient supply towards milk production in early lactation and this demand takes precedence over the provision of optimal conditions for reproduction. In this study we used the bovine Affymetrix 23,000 gene microarray, which contains the most comprehensive set of bovine genes to be assembled and provides a means of investigating the modifying influences of energy balance on liver gene expression. Cows in severe negative energy balance (SNEB) in early lactation showed altered hepatic gene expression in metabolic processes as well as a down regulation of the insulin-like growth factor (IGF) system, where insulin like growth factor-1 (IGF-1), growth hormone receptor variant 1A (GHR1A) and insulin-like growth factor binding protein-acid labile subunit (IGFBP-ALS) were down regulated compared to the cows in the moderate negative energy balance MNEB group, consistent with a five-fold reduction in systemic concentrations of IGF1 in the SNEB group.Cows in SNEB showed elevated expression of key genes involved in the inflammatory response such as interleukin-8 (IL-8). There was a down regulation of genes involved in cellular growth in SNEB cows and moreover a negative regulator of cellular proliferation (HGFIN) was up regulated in SNEB cows, which is likely to compromise adaptation and recovery from NEB. The puma method of analysis revealed that 417 genes were differentially regulated by EB (P

End of Project Report Embryo transfer is being increasingly used in the cattle industry. As well as direct embryo transfers, many embryo-based biotechnologies have the potential to improve cattle production efficiency through enhanced breeding strategies, by facilitating the introduction of desirable traits such as disease resistance and through the production of desirable medical or pharmaceutical products in the milk. These biotechnologies are, however, dependent on a supply of viable in vitro produced (IVP) embryos. While the in vitro fertilization rate is high (80%) in cattle, only about 30 transferable embryos, or blastocysts, are produced from every 100 fertilized oocytes. A major factor affecting the viability of IVP embryos is their failure, in a high proportion of cases, to undergo normal development to the blastocyst stage in the manner of in vivo embryos. The major problem relates to a failure of the cells of IVP embryos to form a compact cell mass when they are 5 - 6 days old. This ultimately leads to developmental problems and compromised viability. Cell compaction is recognized as a critical event in early embryo development and has been associated with marked changes in protein synthesis and phosphorylation in the embryos of some species. This report is the first, to our knowledge, to describe the rate and pattern of protein synthesis and phosphorylation before, during and after compaction in both in vivo and in IVP cattle embryos. The main results are summarised below. European Union 4th Framework Programmes (Contract CT-95-0032)

End of Project Report A major problem for the cattle breeding industry is the high rate of early embryo loss which compromises reproductive efficiency and genetic improvement, resulting in serious financial loss to farmers. An important part of the Teagasc research programme in this area is the investigation of basic parameters of cattle embryo growth, development and viability during the critical period when most of the embryo loss occurs. We have now characterised this period of embryo development and to our knowledge, this is the first report describing the morphology, growth rate, protein content and metabolic activity of cattle embryos during this period. The main results are summarised here and detailed results have been published in the papers listed at the end of this report. Embryo growth rate and protein content increased exponentially between days 8 and 13 after fertilisation. Furthermore, there was a high rate of protein synthetic activity, energy and amino acid metabolism and signal transduction activity, all reaching a peak between days 8 and 13 after fertilisation. Because of the high rate of metabolic activity evident during this time it is likely that the embryos are very susceptible to environmental changes that have the potential to interfere with normal developmental mechanisms. The results arising from this project suggest that the critical period of early embryo loss in cattle may now be narrowed to a time window of day 8 to 13 rather than day 8 to 16 as presumed up to now. The main results are summarised. European Union 4th Framework Programme (Contract CT-95-0190).