title: Endogenous folates and single-carbon metabolism in the ovarian follicle, oocyte and pre-implantation embryo.
abstract: Maternal B-vitamin status at conception can affect fertility and the health of offspring. This study details transcript expression for genes encoding key enzymes in the linked methionine/folate cycles in the bovine oocyte, somatic cells of the ovarian follicle and pre-implantation embryo. Transcripts for all 12 enzymes that were studied and for the two folate receptors (FOLR1 and FOLR2) and reduced folate carrier (SLC19A1) were expressed in liver cells, but transcripts for betaine-homocysteine methyltransferase and methionine adenosyl transferase 1A were absent in all ovarian cells, and transcripts for FOLR2 were absent in embryonic cells. Transcripts for glycine methyltransferase were also absent/weak in cumulus and granulosa cells. The absence of these enzymes could have a profound effect on single-carbon metabolism within the ovary and pre-implantation embryo. Immunocytochemical analysis revealed SLC19A1 protein expression on the plasma and basal-lateral membranes of the pre-implantation embryo. The folate antagonist methotrexate (MTX) enters the cell via SLC19A1, and in the current study, MTX inclusion in bovine/ovine culture media at either 1 or 10 microM from the 1-cell stage inhibited embryo development beyond the 8-cell stage. Hypoxanthine and thymidine (100 microM) increased the proportion of embryos that developed to blastocysts, but the cell number was reduced by 20%. The reduced uptake of [(35)S] methionine into intra-cellular S-adenosylmethionine and S-adenosylhomocysteine pools, together with reduced uptake of glutamate and tryptophan, was consistent with depleted intra-cellular pools of reduced folates. These data provide an insight into the importance of maternal dietary folate/B-vitamin status during the peri-conceptional period.
authors.affiliation: School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE12 5RD, UK.
authors.forename: W Y
authors.initials: WY
authors.lastname: Kwong
authors.forename: S J
authors.initials: SJ
authors.lastname: Adamiak
authors.forename: A
authors.initials: A
authors.lastname: Gwynn
authors.forename: R
authors.initials: R
authors.lastname: Singh
authors.forename: K D
authors.initials: KD
authors.lastname: Sinclair
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000378:metabolism*; D050260:Carbohydrate Metabolism / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D002244:Carbon / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D005260:Female; D005492:Folic Acid / Q000378:metabolism*; D005493:Folic Acid Antagonists / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D053858:Metabolic Networks and Pathways / Q000187:drug effects / Q000235:genetics; D008715:Methionine / Q000378:metabolism; D008727:Methotrexate / Q000494:pharmacology; D008954:Models, Biological; D009865:Oocytes / Q000378:metabolism*; D006080:Ovarian Follicle / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D012756:Sheep
chemical_list: D005493:Folic Acid Antagonists; D002244:Carbon; D005492:Folic Acid; D008715:Methionine; D008727:Methotrexate