caa a22 4500 606235183 CHVBK 20210128101241.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s11010-014-2222-0 doi (NATIONALLICENCE)springer-10.1007/s11010-014-2222-0 Raltitrexed's effect on the development of neural tube defects in mice is associated with DNA damage, apoptosis, and proliferation [Elektronische Daten] [Yanting Dong, Xiuwei Wang, Jianlin Zhang, Zhen Guan, Lin Xu, Jianhua Wang, Ting Zhang, Bo Niu] 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 γ-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 11.5mg/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 γ-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. Springer Science+Business Media New York, 2014 Neural tube defects nationallicence Thymidylate synthase nationallicence Raltitrexed nationallicence DNA damage nationallicence Apoptosis nationallicence Proliferation nationallicence Dong Yanting Department of Biochemistry and Molecular Biology, Shanxi Medical University, 030001, Taiyuan, China aut Wang Xiuwei Capital Institute of Pediatrics, 100020, Beijing, China aut Zhang Jianlin Department of Biochemistry and Molecular Biology, Shanxi Medical University, 030001, Taiyuan, China aut Guan Zhen Capital Institute of Pediatrics, 100020, Beijing, China aut Xu Lin Capital Institute of Pediatrics, 100020, Beijing, China aut Wang Jianhua Capital Institute of Pediatrics, 100020, Beijing, China aut Zhang Ting Capital Institute of Pediatrics, 100020, Beijing, China aut Niu Bo Capital Institute of Pediatrics, 100020, Beijing, China aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 398/1-2(2015-01-01), 223-231 0300-8177 398:1-2<223 2015 398 11010 https://doi.org/10.1007/s11010-014-2222-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-014-2222-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dong Yanting Department of Biochemistry and Molecular Biology, Shanxi Medical University, 030001, Taiyuan, China aut NATIONALLICENCE 700 1- Wang Xiuwei Capital Institute of Pediatrics, 100020, Beijing, China aut NATIONALLICENCE 700 1- Zhang Jianlin Department of Biochemistry and Molecular Biology, Shanxi Medical University, 030001, Taiyuan, China aut NATIONALLICENCE 700 1- Guan Zhen Capital Institute of Pediatrics, 100020, Beijing, China aut NATIONALLICENCE 700 1- Xu Lin Capital Institute of Pediatrics, 100020, Beijing, China aut NATIONALLICENCE 700 1- Wang Jianhua Capital Institute of Pediatrics, 100020, Beijing, China aut NATIONALLICENCE 700 1- Zhang Ting Capital Institute of Pediatrics, 100020, Beijing, China aut NATIONALLICENCE 700 1- Niu Bo Capital Institute of Pediatrics, 100020, Beijing, China aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 398/1-2(2015-01-01), 223-231 0300-8177 398:1-2<223 2015 398 11010