IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro
Follicle selection, a crucial step in maintaining continuous egg production in chickens, is a process that relies on granulosa cells (GCs). In this study, we aimed to identify the key genes that are involved in follicle selection from our previous single-cell transcriptomic data. We used a combinati...
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Elsevier
2024-12-01
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| Series: | Poultry Science |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579124009945 |
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| author | Wenhui Zhang Xuejiao Chen Axiu Guo Zongyi Zhao Bo Zhang Fuwei Li Hao Zhang |
| author_facet | Wenhui Zhang Xuejiao Chen Axiu Guo Zongyi Zhao Bo Zhang Fuwei Li Hao Zhang |
| author_sort | Wenhui Zhang |
| collection | DOAJ |
| description | Follicle selection, a crucial step in maintaining continuous egg production in chickens, is a process that relies on granulosa cells (GCs). In this study, we aimed to identify the key genes that are involved in follicle selection from our previous single-cell transcriptomic data. We used a combination of techniques and assays, including quantitative real-time PCR, immunofluorescence, Oil Red O staining, transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), monodansylcadaverine (MDC) assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, cell counting Kit-8 (CCK-8) assay, and 5-ethynyl-2-deoxyuridine (EdU) assay. Multiple indices, such as cell proliferation, cell differentiation, progesterone synthesis, lipid droplet production, total and free cholesterol content, apoptosis, and autophagy, were measured to determine the states of GCs in vitro. The results demonstrated that overexpression of genes related to insulin-like growth factor 2 (IGF2) or IGF-binding protein 4 (IGFBP4) increases intracellular free cholesterol (progesterone precursors) and lipid droplet production, inhibits apoptosis through increased autophagy, and inhibits cell proliferation. This indicates that IGF2 or IGFBP4 can maintain the survival state and improve differentiation tendency of chicken granulosa cells in vitro. Therefore, this study provides new evidence on the functions of IGFs and IGFBPs in chickens, establishing a crucial experimental foundation for understanding the regulatory mechanisms of follicle selection. In addition, our study contributes to understanding follicular development and improves the egg-laying performance of chickens. |
| format | Article |
| id | doaj-art-ba1d78faf75d493ea235fa53fede70c8 |
| institution | OA Journals |
| issn | 0032-5791 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj-art-ba1d78faf75d493ea235fa53fede70c82025-08-20T01:55:42ZengElsevierPoultry Science0032-57912024-12-011031210441610.1016/j.psj.2024.104416IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitroWenhui Zhang0Xuejiao Chen1Axiu Guo2Zongyi Zhao3Bo Zhang4Fuwei Li5Hao Zhang6State Key Laboratory of Animal Biotech Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR ChinaState Key Laboratory of Animal Biotech Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR ChinaState Key Laboratory of Animal Biotech Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR ChinaState Key Laboratory of Animal Biotech Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR ChinaState Key Laboratory of Animal Biotech Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR ChinaPoultry Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, PR ChinaState Key Laboratory of Animal Biotech Breeding, Beijing Key Laboratory for Animal Genetic Improvement, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China; Corresponding author at: College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.Follicle selection, a crucial step in maintaining continuous egg production in chickens, is a process that relies on granulosa cells (GCs). In this study, we aimed to identify the key genes that are involved in follicle selection from our previous single-cell transcriptomic data. We used a combination of techniques and assays, including quantitative real-time PCR, immunofluorescence, Oil Red O staining, transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), monodansylcadaverine (MDC) assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, cell counting Kit-8 (CCK-8) assay, and 5-ethynyl-2-deoxyuridine (EdU) assay. Multiple indices, such as cell proliferation, cell differentiation, progesterone synthesis, lipid droplet production, total and free cholesterol content, apoptosis, and autophagy, were measured to determine the states of GCs in vitro. The results demonstrated that overexpression of genes related to insulin-like growth factor 2 (IGF2) or IGF-binding protein 4 (IGFBP4) increases intracellular free cholesterol (progesterone precursors) and lipid droplet production, inhibits apoptosis through increased autophagy, and inhibits cell proliferation. This indicates that IGF2 or IGFBP4 can maintain the survival state and improve differentiation tendency of chicken granulosa cells in vitro. Therefore, this study provides new evidence on the functions of IGFs and IGFBPs in chickens, establishing a crucial experimental foundation for understanding the regulatory mechanisms of follicle selection. In addition, our study contributes to understanding follicular development and improves the egg-laying performance of chickens.http://www.sciencedirect.com/science/article/pii/S0032579124009945ChickenFollicle selectionSingle-cell transcriptomeIGF2IGFBP4 |
| spellingShingle | Wenhui Zhang Xuejiao Chen Axiu Guo Zongyi Zhao Bo Zhang Fuwei Li Hao Zhang IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro Poultry Science Chicken Follicle selection Single-cell transcriptome IGF2 IGFBP4 |
| title | IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro |
| title_full | IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro |
| title_fullStr | IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro |
| title_full_unstemmed | IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro |
| title_short | IGF2/IGFBP4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro |
| title_sort | igf2 igfbp4 reduces apoptosis and increases free cholesterol of chicken granulosa cells in vitro |
| topic | Chicken Follicle selection Single-cell transcriptome IGF2 IGFBP4 |
| url | http://www.sciencedirect.com/science/article/pii/S0032579124009945 |
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