PUBLICATION
Knocking down 10-formyltetrahydrofolate dehydrogenase increased oxidative stress and impeded zebrafish embryogenesis by obstructing morphogenetic movement
- Authors
- Chang, W.N., Lee, G.H., Kao, T.T., Lin, C.Y., Hsiao, T.H., Tsai, J.N., Chen, B.H., Chen, Y.H., Wu, H.R., Tsai, H.J., Fu, T.F.
- ID
- ZDB-PUB-140513-121
- Date
- 2014
- Source
- Biochimica et biophysica acta. General subjects 1840(7): 2340-2350 (Journal)
- Registered Authors
- Chen, Yau-Hung
- Keywords
- Cell migration, FDH, Folate metabolism, ROS, Zebrafish
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Embryonic Development/genetics*
- Folic Acid/genetics
- Folic Acid/metabolism*
- Gene Expression Regulation, Developmental
- Gene Knockdown Techniques
- Humans
- Morphogenesis/genetics*
- Morpholinos
- Oxidative Stress/genetics*
- Oxidoreductases Acting on CH-NH Group Donors/genetics*
- Zebrafish/genetics
- Zebrafish/growth & development
- PubMed
- 24747731 Full text @ BBA General Subjects
Citation
Chang, W.N., Lee, G.H., Kao, T.T., Lin, C.Y., Hsiao, T.H., Tsai, J.N., Chen, B.H., Chen, Y.H., Wu, H.R., Tsai, H.J., Fu, T.F. (2014) Knocking down 10-formyltetrahydrofolate dehydrogenase increased oxidative stress and impeded zebrafish embryogenesis by obstructing morphogenetic movement. Biochimica et biophysica acta. General subjects. 1840(7):2340-2350.
Abstract
Background Folate is an essential nutrient for cell survival and embryogenesis. 10-Formyltetrahydrofolate dehydrogenase (FDH) is the most abundant folate enzyme in folate-mediated one-carbon metabolism. 10-Formyltetrahydrofolate dehydrogenase converts 10-formyltetrahydrofolate to tetrahydrofolate and CO2, the only pathway responsible for formate oxidation in methanol intoxication. 10-Formyltetrahydrofolate dehydrogenase has been considered a potential chemotherapeutic target because it was down-regulated in cancer cells. However, the normal physiological significance of 10-Formyltetrahydrofolate dehydrogenase is not completely understood, hampering the development of therapeutic drug/regimen targeting 10-Formyltetrahydrofolate dehydrogenase.
Methods 10-Formyltetrahydrofolate dehydrogenase expression in zebrafish embryos was knocked-down using morpholino oligonucleotides. The morphological and biochemical characteristics of fdh morphants were examined using specific dye staining and whole-mount in-situ hybridization. Embryonic folate contents were determined by HPLC.
Results The expression of 10-formyltetrahydrofolate dehydrogenase was consistent in whole embryos during early embryogenesis and became tissue-specific in later stages. Knocking-down fdh impeded morphogenetic movement and caused incorrect cardiac positioning, defective hematopoiesis, notochordmalformation and ultimate death of morphants. Obstructed F-actin polymerization and delayed epiboly were observed in fdh morphants. These abnormalities were reversed either by adding tetrahydrofolate or antioxidant or by co-injecting the mRNA encoding 10-formyltetrahydrofolate dehydrogenase N-terminal domain, supporting the anti-oxidative activity of 10-formyltetrahydrofolate dehydrogenase and the in vivo function of tetrahydrofolate conservation for 10-formyltetrahydrofolate dehydrogenase N-terminal domain.
Conclusions 10-Formyltetrahydrofolate dehydrogenase functioned in conserving the unstable tetrahydrofolate and contributing to the intracellular anti-oxidative capacity of embryos, which was crucial in promoting proper cell migration during embryogenesis.
General significance These newly reported tetrahydrofolate conserving and anti-oxidative activities of 10-formyltetrahydrofolate dehydrogenase shall be important for unraveling 10-formyltetrahydrofolate dehydrogenase biological significance and the drug development targeting 10-formyltetrahydrofolate dehydrogenase.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping