Uterine Extracellular Vesicles Can Emulate the Long‐Term Effects of Post‐Partum Negative Energy Balance in Dairy Cows

Dairy cows often experience a period of negative energy balance (NEB) during the post-calving period, which can significantly impact economic outcomes due to extended calving-to-conception intervals and overall reduced fertility. This reduction is due, in part, to the impact on uterine biology by high nonesterified fatty acids (NEFA) and beta-hydroxybutyrate concentration. The uterine fluid (UF) contains small extracellular vesicles (UF-EVs) that, through their cargo, including microRNAs (miRNAs), respond to metabolic stress, affecting the uterine environment. This study aimed to assess the long-term impact of NEB intensity on the uterine environment of dairy cows. Post-partum dairy cows were classified based on NEFA concentrations in their blood during the 3 weeks post-calving as having either Low or High NEB. At 30 and 60 DPC, the synchronization protocol was started, and UF samples were collected (corresponding to ~15 days after initiation of the synchronization protocol) to isolate UF-EVs and uterine epithelial cells for miRNA and transcriptome profiling. We also investigated whether UF-EVs could modulate epithelial uterine naïve cells. Our results indicate that the uterine environment of dairy cows experiencing a High NEB post-calving is unfavorable for embryo development at 60-day post-calving. Importantly, we show that UF-EVs can reproduce this phenotype in epithelial uterine naïve cells, suggesting that UF-EVs may act as modulators of the uterine response to metabolic challenges.