1. Academic Validation
  2. The effect of inhibiting glycinamide ribonucleotide formyl transferase on the development of neural tube in mice

The effect of inhibiting glycinamide ribonucleotide formyl transferase on the development of neural tube in mice

  • Nutr Metab (Lond). 2016 Aug 23;13(1):56. doi: 10.1186/s12986-016-0114-x.
Lin Xu  # 1 Li Wang  # 2 JianHua Wang 1 ZhiQiang Zhu 1 Ge Chang 1 Ying Guo 2 XinLi Tian 3 Bo Niu 1
Affiliations

Affiliations

  • 1 Department of Biotechnology, Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, 100020 People's Republic of China.
  • 2 Department of Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan, 030001 People's Republic of China.
  • 3 Department of Cardiovascular Disease, Chinese PLA General Hospital of Beijing Military Region, Beijing, 100020 People's Republic of China.
  • # Contributed equally.
Abstract

Background: Folate deficiency is closely related to the development of neural tube defects (NTDs). However, the exact mechanism is not completely understood. This study aims to induce murine NTDs by inhibiting one of the folate metabolic pathways, de novo purine synthesis and preliminarily investigate the potential mechanisms. The key Enzyme, glycinamide ribonucleotide formyl transferase (GARFT) was inhibited by a specific inhibitor, lometrexol (DDATHF) in the pregnant mice.

Methods: Pregnant mice were intraperitoneally injected with various doses of DDATHF on gestational day 7.5 and embryos were examined for the presence of NTDs on gestational day 11.5. GARFT activity and levels of ATP, GTP, dATP and dGTP were detected in embryonic brain tissue. Proliferation and Apoptosis was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR), immunohistochemical assay and western blotting.

Results: 40 mg kg(-1) body weight (b/w) of DDATHF caused the highest incidence of NTDs (30.8 %) and therefore was selected as the optimal dose to establish murine NTDs. The GARFT activity and levels of ATP, GTP, dATP and dGTP in embryonic brain tissue were significantly decreased after DDATHF treatment. Furthermore, Levels of proliferation-related genes (Pcna, Foxg1 and Ptch1) were downregulated and apoptosis-related genes (Bax, Casp8 and Casp9) were upregulated. Expression of phosphohistone H3 was significantly decreased while expression of cleaved Caspase-3 was greatly increased.

Conclusions: Results indicate that DDATHF induced murine NTDs by disturbing purine metabolism and further led to abnormal proliferation and Apoptosis.

Keywords

Apoptosis; Lometrexol; Metabolism; Mice; Neural tube defects; Proliferation.

Figures
Products