Volume 33, Issue 9 p. 915-923
Research Article

Influence of the antifolate drug Methotrexate on the development of murine neural tube defects and genomic instability

Jie Zhao

Jie Zhao

Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, 030001 China

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Tao Guan

Tao Guan

Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, 030001 China

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Jianhua Wang

Jianhua Wang

Capital Institute of Pediatrics, Beijing, 100020 China

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Qian Xiang

Qian Xiang

Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100730 China

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Mingsheng Wang

Mingsheng Wang

Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100730 China

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Xiuwei Wang

Xiuwei Wang

Capital Institute of Pediatrics, Beijing, 100020 China

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Zhen Guan

Zhen Guan

Capital Institute of Pediatrics, Beijing, 100020 China

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Qiu Xie

Qiu Xie

Capital Institute of Pediatrics, Beijing, 100020 China

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Bo Niu

Corresponding Author

Bo Niu

Capital Institute of Pediatrics, Beijing, 100020 China

Correspondence to: Ting Zhang, Capital Institute of Pediatrics, Beijing, 100020, China. E-mail: [email protected].

Bo Niu, Capital Institute of Pediatrics, Beijing, 100020, China. E-mail: [email protected]

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Ting Zhang

Corresponding Author

Ting Zhang

Capital Institute of Pediatrics, Beijing, 100020 China

Correspondence to: Ting Zhang, Capital Institute of Pediatrics, Beijing, 100020, China. E-mail: [email protected].

Bo Niu, Capital Institute of Pediatrics, Beijing, 100020, China. E-mail: [email protected]

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First published: 13 July 2012
Citations: 19

Both authors contributed equally to this manuscript.

ABSTRACT

Impaired folate metabolism is considered a risk factor for neural tube defects (NTDs). However, the relationship between folate deficiency and the risk of NTDs remains unclear, because experimentally induced dietary folate deficiency is insufficient to cause NTDs in non-mutant mice. Methotrexate (MTX) is a specific folate antagonist that competitively inhibits dihydrofolate reductase (DHFR) activity. The objective of this study was to develop a folate dysmetabolism murine model, and study the development of NTDs and its mechanism. Pregnant mice were injected with different doses of MTX [0, 0.5, 1.0, 3.0, 4.5 and 6.0 mg kg–1 body weight (b/w) intraperitoneally (i.p.)] on gestational day 7.5 and sacrificed on gestational day 11.5. DHFR activity in embryonic tissues was detected, and folate concentrations were analyzed using LC/MS/MS. Copy number variations (CNVs) in neural tube tissues were detected using array comparative genomic hybridization (aCGH). A dose of MTX 4.5 mg kg–1 b/w, resulted in the highest incidence of NTDs (31.4%) compared with the other groups, and DHFR activities, 5-MeTHF and 5-FoTHF concentrations in embryonic tissues decreased significantly after MTX injection. Furthermore, we found three high-confidence CNVs on chromosome X using aCGH, which was confirmed by RT-PCR and MassARRAY. These results indicate that MTX could cause a folate-associated dysmetabolism, which is similar to that of dietary folate deficiency in mice. The presence of CNVs in neural tube tissues was associated with the development of NTDs. Copyright © 2012 John Wiley & Sons, Ltd.