Precise Dose of Folic Acid Supplementation Is Essential for Embryonic Heart Development in Zebrafish
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Zebrafish Maintenance and Care
2.2. Visible Light Imaging and Fluorescence Imaging of Zebrafish Embryos
2.3. Microinjection of Zebrafish Embryos with CRISPR/Cas9 Knock-Out
2.4. Protein Structure Prediction
2.5. Cardiac Physiology Measurement
2.6. In Situ Hybridization
2.7. Quantitative Real-Time PCR
2.8. Luciferase Assays
2.9. Statistical Analysis
2.10. Behavioral Test
2.11. S-Adenosylmethionine Content Determination
2.12. Assessment of DNA Methylation Level
3. Results
3.1. Folic Acid and MTX Showed Dose-Dependent Effects on Early Cardiac Development in Zebrafish
3.2. Insufficient or Excessive Folic Acid Leads to Heart Looping Defect and Abnormal Homocysteine Metabolism
3.3. Abnormal Folate Metabolism Leads to Behavior Disorder of Zebrafish Embryos
3.4. CRISPR/Cas9 Mediated mthfr Gene Knock-Out Model in Zebrafish
3.5. Folic Acid Deficiency and Excessive Supplement Lead to the Changes of Heart Related Genes
3.6. Folate Metabolism Changes Defect the Methylation of hand2
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Han, X.; Wang, B.; Jin, D.; Liu, K.; Wang, H.; Chen, L.; Zu, Y. Precise Dose of Folic Acid Supplementation Is Essential for Embryonic Heart Development in Zebrafish. Biology 2022, 11, 28. https://doi.org/10.3390/biology11010028
Han X, Wang B, Jin D, Liu K, Wang H, Chen L, Zu Y. Precise Dose of Folic Acid Supplementation Is Essential for Embryonic Heart Development in Zebrafish. Biology. 2022; 11(1):28. https://doi.org/10.3390/biology11010028
Chicago/Turabian StyleHan, Xuhui, Bingqi Wang, Dongxu Jin, Kuang Liu, Hongjie Wang, Liangbiao Chen, and Yao Zu. 2022. "Precise Dose of Folic Acid Supplementation Is Essential for Embryonic Heart Development in Zebrafish" Biology 11, no. 1: 28. https://doi.org/10.3390/biology11010028