Regulation of Follicular Development in Chickens: WIF1 Modulates Granulosa Cell Proliferation and Progesterone Synthesis via Wnt/β-Catenin Signaling Pathway
Abstract
:1. Introduction
2. Results
2.1. Expression Characteristics of WIF1 in Chickens
2.2. WIF1 Inhibits Proliferation of Chicken GCs from Prehierarchal Follicles
2.3. WIF1 Promotes the Apoptosis of Chicken Prehierarchal GCs
2.4. WIF1 Promotes the Secretion of Progesterone with the Participation of FSH in Chicken GCs
2.5. WIF1 Inhibits Wnt Signaling Activity in Chicken GCs
2.6. WIF1 Stimulates Metabolic Activity of Chicken Prehierarchal GCs
2.7. WIF1 Promotes Progesterone Synthesis through Upregulation of ESR2 in GCs
2.8. WIF1 Suppresses TC Proliferation and Promotes Apoptosis
3. Discussion
4. Materials and Methods
4.1. Ethics Statement and Experimental Animals
4.2. Cell Isolation and Culture
4.3. Plasmid Construction, RNA Oligonucleotide Synthesis, and Cell Transfection
4.4. Quantitative Real-Time PCR (qRT-PCR)
4.5. Western Blotting
4.6. 5-Ethynyl-2′-Deoxyuridine (EdU) Assay
4.7. Cell Counting Kit-8 (CCK 8) Assay
4.8. Flow Cytometry
4.9. Enzyme-Linked Immunosorbent Assay (ELISA)
4.10. Dual-Luciferase Reporter Assay
4.11. RNA-seq and Bioinformatics Analysis
4.12. Co-Transfection of Plasmid DNA and Small Interfering RNA (siRNA)
4.13. Immunofluorescence Staining
4.14. TdT-Mediated dUTP Nick-End Labeling (TUNEL) Assay
4.15. Statistical Analysis and Data Visualization
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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Nie, R.; Zhang, W.; Tian, H.; Li, J.; Ling, Y.; Zhang, B.; Zhang, H.; Wu, C. Regulation of Follicular Development in Chickens: WIF1 Modulates Granulosa Cell Proliferation and Progesterone Synthesis via Wnt/β-Catenin Signaling Pathway. Int. J. Mol. Sci. 2024, 25, 1788. https://doi.org/10.3390/ijms25031788
Nie R, Zhang W, Tian H, Li J, Ling Y, Zhang B, Zhang H, Wu C. Regulation of Follicular Development in Chickens: WIF1 Modulates Granulosa Cell Proliferation and Progesterone Synthesis via Wnt/β-Catenin Signaling Pathway. International Journal of Molecular Sciences. 2024; 25(3):1788. https://doi.org/10.3390/ijms25031788
Chicago/Turabian StyleNie, Ruixue, Wenhui Zhang, Haoyu Tian, Junying Li, Yao Ling, Bo Zhang, Hao Zhang, and Changxin Wu. 2024. "Regulation of Follicular Development in Chickens: WIF1 Modulates Granulosa Cell Proliferation and Progesterone Synthesis via Wnt/β-Catenin Signaling Pathway" International Journal of Molecular Sciences 25, no. 3: 1788. https://doi.org/10.3390/ijms25031788