Journal: Molecular Reproduction and Development

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

doi: 10.1002/mrd.70062

Figure Lengend Snippet: Schematic overview of the experimental model. (A) Blood samples were collected from individual cows ( n = 7) at 7‐, 14‐, and 21‐day post‐calving (DPC) to determine the animal's energy status based on NEFA concentration. At 30 DPC, animals were evaluated by metricheck, transrectal ultrasonography, and uterine cytology. Only healthy animals (Low NEB = 3; High NEB = 4) had the estrous cycle synchronized. At 60 DPC the same animals were reassessed for reproductive parameters, and healthy animals (Low NEB = 3; High NEB = 3) had the estrous cycle synchronized. (B) After evaluation, the animals were submitted to an estrus synchronization protocol (PGF: prostaglandin F 2α ; EC: estradiol cypionate, GnRH: gonadotropin‐releasing hormone, AI: artificial insemination) and at 5 days after AI, the uterine fluid collection was performed (Day 0 = day of AI). Extracellular vesicles and epithelial uterine cells were separated from the uterine fluid. (C) To collect epithelial uterine naïve cells, the estrous cycles of dairy heifers were synchronized and 4 days after estrous, luminal epithelial cells from the ipsilateral horn to the corpus luteum were collected in vivo with a cytology brush ( n = 3). After collection, cells were cultured, subcultures were performed, and cells were treated for 6 h with extracellular vesicles from uterine fluid (UF‐EVs). BHB, β‐hydroxybutyrate; NEB, negative energy balance; NEFA, nonesterified fatty acids. Created with BioRender ( https://biorender.com/ ).

Article Snippet: NEFA and BHB concentrations were quantified using the NEFA assay kit (FA115; Randox laboratory limited, United Kingdom) and the Ranbut D‐3‐hydroxybutrate with BHBA kit (RB1007; Randox laboratory limited, United Kingdom), respectively, following the manufacturer's protocols.

Techniques: Concentration Assay, In Vivo, Cell Culture

Journal: Molecular Reproduction and Development

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

doi: 10.1002/mrd.70062

Figure Lengend Snippet: NEFA and BHB concentration in the serum. (A) Blood samples to evaluate NEFA were evaluated once a week during the 3 weeks post‐calving (DPC; p < 0.05). (B) BHB were evaluated once a week during the 3 weeks post‐calving ( p < 0.05). n = 7 total; Low NEB = 3 (blue line); High NEB = 4 (red line). BHB, β‐hydroxybutyrate; NEFA, nonesterified fatty acids.

Article Snippet: NEFA and BHB concentrations were quantified using the NEFA assay kit (FA115; Randox laboratory limited, United Kingdom) and the Ranbut D‐3‐hydroxybutrate with BHBA kit (RB1007; Randox laboratory limited, United Kingdom), respectively, following the manufacturer's protocols.

Techniques: Concentration Assay