m6A Methylation Mediates the Function of the circRNA-08436/miR-195/ELOVL6 Axis in Regards to Lipid Metabolism in Dairy Goat Mammary Glands
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. Experimental Animals and Preparation of Samples
2.3. Differential Expression Analysis of circRNAs
2.4. Cell Culture and Treatment
2.5. Determination of TAG and Cholesterol
2.6. Determination of Intracellular Fatty Acid Content
2.7. Oil Red O Staining
2.8. 5-Ethynyl-2′-Deoxyuridine (EdU) Experiment
2.9. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)
2.10. Construction of Circ08436 Overexpression Vector
2.11. Western Blotting
2.12. Dual-Luciferase Activity Analysis
2.13. Statistical Analysis
3. Results
3.1. High-Throughput Sequencing of circRNAs from Mammary Gland Tissue of Dairy Goats
3.2. Differential Expression Analysis of mRNAs
3.3. Enrichment Analysis and Pathway Network Diagram of Differential Genes
3.4. Adsorption and Binding of circRNA-08436 to miR-195
3.5. Specific Targeting of miR-195 on ELOVL6
3.6. Functional Verification of circRNA-08436 in GMECs
3.7. Functional Role of miR-195 and ELOVL6 in GMECs
3.8. Combination of circ08436 with miR-195 to Relieve the Inhibition of ELOVL6
3.9. Regulation of circRNA-08436 by m6A Methylation
4. Discussion
4.1. CircRNA-08436 Can Promote TAG Synthesis
4.2. CircRNA-08436 Can Competitively Bind to miR-195, Thereby Deregulating the Inhibitory Effect of miR-195 on the Target Gene ELOVL6
4.3. YTHDC2 May Play a Role in the Degradation of circRNA-08436 in the Cytoplasm
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Fatty Acid | Control | circ08436 |
---|---|---|
C16:0 (%) | 21.41 ± 0.14 | 41.12 ± 0.56 ** |
C16:1 (%) | 10.12 ± 0.57 | 2.10 ± 0.34 ** |
C18:0 (%) | 10.57 ± 0.34 | 18.41 ± 0.25 * |
C18:1 (%) | 48.14 ± 0.35 | 34.83 ± 0.41 ** |
C18:2 (%) | 9.76 ± 0.24 | 3.54 ± 0.24 * |
SFA (%) | 31.98 | 59.53 |
UFA (%) | 68.02 | 40.47 |
UFA/SFA | 2.13 | 0.68 |
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Wang, Y.; Wu, Y.; Yang, S.; Gao, R.; Lv, X.; Yang, Z.; Jiao, P.; Zhang, N.; Loor, J.J.; Chen, Z. m6A Methylation Mediates the Function of the circRNA-08436/miR-195/ELOVL6 Axis in Regards to Lipid Metabolism in Dairy Goat Mammary Glands. Animals 2024, 14, 1715. https://doi.org/10.3390/ani14121715
Wang Y, Wu Y, Yang S, Gao R, Lv X, Yang Z, Jiao P, Zhang N, Loor JJ, Chen Z. m6A Methylation Mediates the Function of the circRNA-08436/miR-195/ELOVL6 Axis in Regards to Lipid Metabolism in Dairy Goat Mammary Glands. Animals. 2024; 14(12):1715. https://doi.org/10.3390/ani14121715
Chicago/Turabian StyleWang, Yu, Yanni Wu, Sitian Yang, Rui Gao, Xiaoyang Lv, Zhangping Yang, Peixin Jiao, Ning Zhang, Juan J. Loor, and Zhi Chen. 2024. "m6A Methylation Mediates the Function of the circRNA-08436/miR-195/ELOVL6 Axis in Regards to Lipid Metabolism in Dairy Goat Mammary Glands" Animals 14, no. 12: 1715. https://doi.org/10.3390/ani14121715