Abstract
Highly proliferative cells rely on one carbon (1C) metabolism for production of formate required for synthesis of purines and thymidine for nucleic acid synthesis. This study was to determine if extracellular serine and/or glucose and fructose contribute the production of formate in ovine conceptuses. Suffolk ewes (n = 8) were synchronized to estrus, bred to fertile rams, and conceptuses were collected on Day 17 of gestation. Conceptuses were either snap frozen in liquid nitrogen (n = 3) or placed in culture in medium (n = 5) containing either: 1) 4 mM D-glucose + 2 mM [U-13C]serine; 2) 6 mM glycine + 4 mM D-glucose + 2 mM [U-13C]serine; 3) 4 mM D-fructose + 2 mM [U-13C]serine; 4) 6 mM glycine + 4 mM D-fructose + 2 mM [U-13C]serine; 5) 4 mM D-glucose + 4 mM D-fructose + 2 mM [U-13C]serine; or 6) 6 mM glycine + 4 mM D-glucose + 4 mM D-fructose + 2 mM [U-13C]serine. After 2 h incubation, conceptuses in their respective culture medium were homogenized and the supernatant analyzed for 12C- and 13C-formate by gas chromatography and amino acids by high performance liquid chromatography. Ovine conceptuses produced both 13C- and 12C-formate, indicating that the [U-13C]serine, glucose, and fructose were utilized to generate formate, respectively. Greater amounts of 12C-formate than 13C-formate were produced, indicating that the ovine conceptus utilized more glucose and fructose than serine to produce formate. This study is the first to demonstrate that both 1C metabolism and serinogenesis are active metabolic pathways in ovine conceptuses during the peri-implantation period of pregnancy, and that hexose sugars are the preferred substrate for generating formate required for nucleotide synthesis for proliferating trophectoderm cells.
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Acknowledgements
This research was supported by Agriculture and Food Research Initiative Competitive Grant No. 2018-67015-28093 from the USDA National Institute of Food and Agriculture. The authors sincerely thank those involved in animal care and technical support for the generation and collection of these samples, as well as the execution of experiments and data analysis.
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726_2022_3212_MOESM1_ESM.tif
Supplementary file1 Expression of mRNAs required for nutrient transport in conceptus tissue on Day 17 of gestation after a 2 h incubation with [U-13C]serine. Data are expressed as relative expression of mRNAs, means ± SEM. n = 3 samples per group (TIF 5264 kb)
726_2022_3212_MOESM2_ESM.tif
Supplementary file2 Expression of mRNAs required for serinogenesis in conceptus tissue from Day 17 of gestation after a 2 h incubation with [U-13C]serine. Data are expressed as relative expression of mRNAs, means ± SEM. n = 3 samples per group (TIF 3759 kb)
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Halloran, K.M., Stenhouse, C., Moses, R.M. et al. The ovine conceptus utilizes extracellular serine, glucose, and fructose to generate formate via the one carbon metabolism pathway. Amino Acids 55, 125–137 (2023). https://doi.org/10.1007/s00726-022-03212-x
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DOI: https://doi.org/10.1007/s00726-022-03212-x