1. MicroRNA-375 overexpression disrupts cardiac development of Zebrafish (Danio rerio) by targeting notch2
- Author
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Jingai Zhu, Yanrong Fu, Meiling Tong, Sisi Zhuang, Mengmeng Li, Jingyun Li, and Xiaoshan Hu
- Subjects
0106 biological sciences ,0301 basic medicine ,Danio ,Notch signaling pathway ,Embryonic Development ,Plant Science ,In situ hybridization ,01 natural sciences ,03 medical and health sciences ,Downregulation and upregulation ,Mir-375 ,microRNA ,Animals ,Luciferase ,Receptor, Notch2 ,Zebrafish ,biology ,Heart ,Cell Biology ,General Medicine ,biology.organism_classification ,Cell biology ,MicroRNAs ,030104 developmental biology ,010606 plant biology & botany - Abstract
MicroRNAs are small noncoding RNAs that are important for proper cardiac development. In our previous study of fetuses with ventricular septal defects, we discovered that microRNA-375 (miR-375) is obviously upregulated compared with that in healthy controls. Our study also confirmed that miR-375 is crucial for cardiomyocyte differentiation. This research mainly focused on the biological significance and mechanism of miR-375 using a zebrafish model. We injected zebrafish embryos with 1-2 nl of a miR-375 mimic at various concentrations (0/2/4/8 μM) or with negative control. The deformation and mortality rates were separately assessed. The different expression levels of miR-375 and related genes were examined by qRT-PCR, and luciferase assays and in situ hybridization were used to clarify the mechanism of miR-375 during embryonic development. Overexpression of miR-375 disrupted the cardiac development of zebrafish embryos. Disruption of miR-375 led to a decreased heart rate, pericardial edema, and abnormal cardiac looping. Various genes involved in cardiac development were downregulated due to the overexpression of miR-375. Moreover, the NOTCH signaling pathway was affected, and the luciferase reporter gene assays confirmed notch2, which was predicted by bioinformatics analysis, as the target gene of miR-375. Our findings demonstrated that the overexpression of miR-375 is detrimental to embryonic development, including cardiac development, and can partially simulate a multisystemic disorder. MiR-375 has an important role during cardiac morphogenesis of zebrafish embryos by targeting notch2, indicating its potential as a diagnostic marker.
- Published
- 2020
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