Strategy of Hepatic Metabolic Defects Induced by beclin1 Heterozygosity in Adult Zebrafish
Abstract
:1. Introduction
2. Results
2.1. CRISPR/Cas9-Mediated Targeted Mutagenesis of atg7 and beclin1 in Zebrafish
2.2. Beclin1 Heterozygosity Affects Liver Histology and Causes High Mortality Rate in Male Fish
2.3. Heterozygosity of beclin1 Alters Hepatic Energy Metabolism in Male Zebrafish
2.4. Beclin1+/− Affects Hepatic Glucose/Lipid Flux on the Genetic Level
2.5. Unlike atg7+/−, Heterozygosity of beclin1 Suppresses Autophagy via Activating PI3K/AKT Pathway
2.6. Induction of IL6/STAT3 Inflammatory Pathway and Downregulation of Apoptosis Pathway in beclin1+/− Liver
2.7. Heterozygosity of beclin1 Enhances Hepatocellular Proliferation and Inflammation Probably Due to Apoptosis Suppression
3. Discussion
4. Materials and Methods
4.1. Establishment of atg7 and beclin1 Mutant Lines and Calculation of Survival Rates
4.2. Assessments of Liver Histology
4.3. RNA Extraction and Quantitative Real-Time PCR (qRT-PCR)
4.4. Protein Extraction and Western Blotting
4.5. Immunohistochemistry, TUNEL Assay, and Immunofluorescence
4.6. Image Acquisition and Digital Image Analysis
4.7. Transcriptomic Sequencing (RNA-Seq) and Gene Ontology
4.8. Statistical Analysis and Graphs Preparation
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Mawed, S.A.; He, Y.; Zhang, J.; Mei, J. Strategy of Hepatic Metabolic Defects Induced by beclin1 Heterozygosity in Adult Zebrafish. Int. J. Mol. Sci. 2020, 21, 1533. https://doi.org/10.3390/ijms21041533
Mawed SA, He Y, Zhang J, Mei J. Strategy of Hepatic Metabolic Defects Induced by beclin1 Heterozygosity in Adult Zebrafish. International Journal of Molecular Sciences. 2020; 21(4):1533. https://doi.org/10.3390/ijms21041533
Chicago/Turabian StyleMawed, Suzan Attia, Yan He, Jin Zhang, and Jie Mei. 2020. "Strategy of Hepatic Metabolic Defects Induced by beclin1 Heterozygosity in Adult Zebrafish" International Journal of Molecular Sciences 21, no. 4: 1533. https://doi.org/10.3390/ijms21041533