67 Influence of Metabolic Status and Genetic Merit for Fertility on Proteomic Composition of Bovine Uterine Luminal Fluid

Abstract

Intensive selection strategies focusing on increased milk yield over several decades has been associated with a decline in fertility in dairy cows. To study the effect of the genetic merit for fertility and the metabolic status of the female on the oocyte, early embryo and the maternal environment, 2 animal models were established. The genetic merit model involved Holstein heifers with a low (LFH) and high fertility (HFH) index and heifers from the Montbéliarde breed (MBD), known to have good reproductive performance. The metabolic model comprised samples from maiden heifers (MH), postpartum lactating cows (Lact), and non-lactating cows (dried off immediately after calving; Dry). A common pool of Day 7 embryos recovered from superovulated and artificially inseminated Holstein heifers were transferred into synchronised recipients (1/recipient) of the above-mentioned animal models and uterine lumen fluid (ULF) of confirmed pregnant animals was recovered on Day 19 post-oestrus. As communication between the conceptus and the uterine environment is crucial for the successful establishment of pregnancy, we analysed uterine luminal fluid of pregnant cows from both models using a holistic proteomic approach. Using nano-liquid chromatography-tandem mass spectrometry analysis combined with a label-free quantification approach, we analysed the uterine luminal fluid from the uterine horn ipsilateral to the corpus luteum (where the conceptus was located in all cases). A total of 2127 proteins were quantified in all samples of both models. Among this set of proteins, 458 were found to differ significantly (P-value < 0.05) in abundance between the groups of the genetic model, and 141 were altered in abundance in the metabolic model. The majority of proteome differences in ULF samples was found comparing HFH to the LFH group (358) and between Dry and Lact cows (70) in the metabolic model. Evaluation of this dataset using bioinformatic tools comprising DAVID GO and gene set enrichment analysis revealed that the affected proteins were predominantly assigned to the terms “translation”, “monosaccharide metabolic process”, “enzyme inhibitor activity”, “lipid binding”, and “response to oxidative stress”. Our study revealed that metabolic status and genetic merit for fertility lead to quantitative molecular differences at the level of proteins in uterine fluid of pregnant animals, thus altering the microenvironment for the early conceptus.

This research was funded by European Union Seventh Framework Programme FP7/2007-2013 under grant agreement no. 312097 (‘FECUND’).