Metabolomic profiling of neat uterine luminal fluid in cows: local enrichment and nutritional modulation

Uterine luminal fluid influences embryonic development and the subsequent phenotype of offspring, yet its detailed metabolomic composition remains poorly characterized. Here, minimally invasive transcervical techniques were employed to collect neat uterine fluid from postpartum dairy cows and cyclic beef cows to allow for metabolomic profiling via targeted mass spectrometry. Objectives were to 1) compare the metabolomic profile of uterine fluid with plasma in dairy cows and 2) assess the impact of dietary rumen-protected methionine and stage of estrous cycle (day 0 vs 7) on plasma and uterine fluid metabolomic profile in beef cows. Results revealed that the concentrations of many metabolites, including amino acids, signaling molecules (e.g. dopamine, gamma-aminobutyric acid) and lipids (e.g. ceramides, diacylglycerols), were higher in uterine fluid than in plasma. An oral bolus of rumen-protected methionine increased uterine concentration of methionine on day 0 of the estrous cycle. The uterine metabolome remained relatively stable between days 0 and 7 although there was temporal variability for a select number of metabolites (cysteine, methionine, methionine sulfoxide, asymmetric dimethylarginine, ceramides, and glycerophospholipids). Correlations between plasma and uterine fluid concentrations were strong or moderate for many amino acids. Collectively, these findings highlight that the uterine lumen is a specialized, selectively regulated biochemical compartment.