PMID- 25245820 OWN - NLM STAT- MEDLINE DCOM- 20160115 LR - 20211021 IS - 1573-4919 (Electronic) IS - 0300-8177 (Linking) VI - 398 IP - 1-2 DP - 2015 Jan TI - Raltitrexed's effect on the development of neural tube defects in mice is associated with DNA damage, apoptosis, and proliferation. PG - 223-31 LID - 10.1007/s11010-014-2222-0 [doi] AB - The causal metabolic pathway and the underlying mechanism between folate deficiency and neural tube defects (NTDs) remain obscure. Thymidylate (dTMP) is catalyzed by thymidylate synthase (TS) using the folate-derived one-carbon unit as the sole methyl donor. This study aims to examine the role of dTMP biosynthesis in the development of neural tube in mice by inhibition of TS via a specific inhibitor, raltitrexed (RTX). Pregnant mice were intraperitoneally injected with various doses of RTX on gestational day 7.5, and embryos were examined for the presence of NTDs on gestational day 11.5. TS activity and changes of dUMP and dTMP levels were measured following RTX treatment at the optimal dose. DNA damage was determined by detection of phosphorylated replication protein A2 (RPA2) and gamma-H2AX in embryos with NTDs induced by RTX. Besides, apoptosis and proliferation were also analyzed in RTX-treated embryos with NTDs. We found that NTDs were highly occurred by the treatment of RTX at the optimal dose of 11.5 mg/kg b/w. RTX treatment significantly inhibited TS activity. Meanwhile, dTMP was decreased associated with the accumulation of dUMP in RTX-treated embryos. Phosphorylated RPA2 and gamma-H2AX were significantly increased in RTX-treated embryos with NTDs compared to control. More apoptosis and decreased proliferation were also found in embryos with NTDs induced by RTX. These results indicate that impairment of dTMP biosynthesis caused by RTX led to the development of NTDs in mice. DNA damage and imbalance between apoptosis and proliferation may be potential mechanisms. FAU - Dong, Yanting AU - Dong Y AD - Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, 030001, China. FAU - Wang, Xiuwei AU - Wang X FAU - Zhang, Jianlin AU - Zhang J FAU - Guan, Zhen AU - Guan Z FAU - Xu, Lin AU - Xu L FAU - Wang, Jianhua AU - Wang J FAU - Zhang, Ting AU - Zhang T FAU - Niu, Bo AU - Niu B LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140923 PL - Netherlands TA - Mol Cell Biochem JT - Molecular and cellular biochemistry JID - 0364456 RN - 0 (Deoxyuracil Nucleotides) RN - 0 (Folic Acid Antagonists) RN - 0 (Histones) RN - 0 (Protein Subunits) RN - 0 (Quinazolines) RN - 0 (Replication Protein A) RN - 0 (Thiophenes) RN - 0 (gamma-H2AX protein, mouse) RN - 365-07-1 (Thymidine Monophosphate) RN - 964-26-1 (2'-deoxyuridylic acid) RN - EC 2.1.1.45 (Thymidylate Synthase) RN - FCB9EGG971 (raltitrexed) SB - IM MH - Animals MH - Apoptosis/*drug effects MH - Blotting, Western MH - Cell Proliferation/*drug effects MH - *DNA Damage MH - Deoxyuracil Nucleotides/metabolism MH - Embryo, Mammalian/*drug effects/metabolism/pathology MH - Female MH - Folic Acid Antagonists/administration & dosage/toxicity MH - Gestational Age MH - Histones/metabolism MH - Injections, Intraperitoneal MH - Male MH - Mice, Inbred C57BL MH - Neural Tube Defects/chemically induced/genetics/*metabolism MH - Phosphorylation/drug effects MH - Pregnancy MH - Protein Subunits/metabolism MH - Quinazolines/administration & dosage/*toxicity MH - Replication Protein A/metabolism MH - Thiophenes/administration & dosage/*toxicity MH - Thymidine Monophosphate/metabolism MH - Thymidylate Synthase/antagonists & inhibitors/metabolism EDAT- 2014/09/24 06:00 MHDA- 2016/01/16 06:00 CRDT- 2014/09/24 06:00 PHST- 2014/06/11 00:00 [received] PHST- 2014/09/15 00:00 [accepted] PHST- 2014/09/24 06:00 [entrez] PHST- 2014/09/24 06:00 [pubmed] PHST- 2016/01/16 06:00 [medline] AID - 10.1007/s11010-014-2222-0 [doi] PST - ppublish SO - Mol Cell Biochem. 2015 Jan;398(1-2):223-31. doi: 10.1007/s11010-014-2222-0. Epub 2014 Sep 23.