Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload
Abstract Background Follicle development is a complicated biological process that produces mature oocytes, and requires nutrients, growth factors, and steroids produced by ovarian granulosa cells (GCs). High fork head box J2 (FOXJ2) expression might negatively regulate ovarian function; however, the...
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BMC
2025-04-01
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| Series: | Journal of Ovarian Research |
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| Online Access: | https://doi.org/10.1186/s13048-025-01651-0 |
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| author | Yunxia Zhang Qiqian Wu Furong Bai Yanqin Hu Bufang Xu Yujie Tang Jingwen Wu |
| author_facet | Yunxia Zhang Qiqian Wu Furong Bai Yanqin Hu Bufang Xu Yujie Tang Jingwen Wu |
| author_sort | Yunxia Zhang |
| collection | DOAJ |
| description | Abstract Background Follicle development is a complicated biological process that produces mature oocytes, and requires nutrients, growth factors, and steroids produced by ovarian granulosa cells (GCs). High fork head box J2 (FOXJ2) expression might negatively regulate ovarian function; however, the mechanism is unclear. This study aimed to investigate the effect and mechanism of FOXJ2 overexpression in GCs on regulating follicle development and fertility. Methods A GC-specific conditional Foxj2 knock-in mouse model (Amh-cre; Foxj2 tg/tg mouse) was generated. Reproductive phenotypes were compared between Amh-cre; Foxj2 tg/tg and control mice using fertility evaluation, oocyte collection, estrus cycle analysis, hormone evaluation, and ovarian follicle assessment. Then, RNA sequencing and bioinformatic analyses were used to detect the altered transcriptome of GCs collected from the Amh-cre; Foxj2 tg/tg and wild-type mice. Western blotting, transmission electron microscopy, immunofluorescence staining, and flow cytometry were used to explore apoptosis and mitochondrial calcium homeostasis. Furthermore, Chromatin immunoprecipitation-PCR and dual-luciferase reporter assays were used to detect the target gene of FOXJ2. Moreover, short hairpin RNA interference was performed on primary GCs and human ovarian granulosa-like tumor (KGN) cells to explore the relationship between FOXJ2 and its target gene in apoptosis and mitochondrial calcium overload. Results FOXJ2 overexpression in GCs led to reduced fertility, hormonal abnormalities, and follicle atresia, starting at the initiation of sexual maturity, resulting in a premature ovarian insufficiency (POI)-like phenotype. Increased apoptosis and mitochondrial calcium overload were detected in the GCs of Amh-cre; Foxj2 tg/tg mice. Mcu (encoding a mitochondrial calcium uniporter) was observed to be upregulated in the GCs of the Amh-cre; Foxj2 tg/tg mice and was a direct target of FOXJ2. Moreover, Mcu knockdown restored mitochondrial calcium homeostasis and reduced the apoptosis in the GCs of the Amh-cre; Foxj2 tg/tg mice and in KGN cells transfected with FOXJ2-overexpression lentivirus. |
| format | Article |
| id | doaj-art-c7b8a80f935c42bfb828e6c308aeeaa1 |
| institution | DOAJ |
| issn | 1757-2215 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
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| series | Journal of Ovarian Research |
| spelling | doaj-art-c7b8a80f935c42bfb828e6c308aeeaa12025-08-20T03:06:52ZengBMCJournal of Ovarian Research1757-22152025-04-0118111710.1186/s13048-025-01651-0Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overloadYunxia Zhang0Qiqian Wu1Furong Bai2Yanqin Hu3Bufang Xu4Yujie Tang5Jingwen Wu6Department of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of MedicineDepartment of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of MedicineShanghai Key Laboratory of Reproductive MedicineDepartment of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of MedicineDepartment of Andrology, Center for Men’s Health, Department of ART, Institute of Urology, Shanghai General Hospital, Urologic Medical Center, Shanghai Jiao Tong University School of MedicineDepartment of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of MedicineDepartment of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of MedicineAbstract Background Follicle development is a complicated biological process that produces mature oocytes, and requires nutrients, growth factors, and steroids produced by ovarian granulosa cells (GCs). High fork head box J2 (FOXJ2) expression might negatively regulate ovarian function; however, the mechanism is unclear. This study aimed to investigate the effect and mechanism of FOXJ2 overexpression in GCs on regulating follicle development and fertility. Methods A GC-specific conditional Foxj2 knock-in mouse model (Amh-cre; Foxj2 tg/tg mouse) was generated. Reproductive phenotypes were compared between Amh-cre; Foxj2 tg/tg and control mice using fertility evaluation, oocyte collection, estrus cycle analysis, hormone evaluation, and ovarian follicle assessment. Then, RNA sequencing and bioinformatic analyses were used to detect the altered transcriptome of GCs collected from the Amh-cre; Foxj2 tg/tg and wild-type mice. Western blotting, transmission electron microscopy, immunofluorescence staining, and flow cytometry were used to explore apoptosis and mitochondrial calcium homeostasis. Furthermore, Chromatin immunoprecipitation-PCR and dual-luciferase reporter assays were used to detect the target gene of FOXJ2. Moreover, short hairpin RNA interference was performed on primary GCs and human ovarian granulosa-like tumor (KGN) cells to explore the relationship between FOXJ2 and its target gene in apoptosis and mitochondrial calcium overload. Results FOXJ2 overexpression in GCs led to reduced fertility, hormonal abnormalities, and follicle atresia, starting at the initiation of sexual maturity, resulting in a premature ovarian insufficiency (POI)-like phenotype. Increased apoptosis and mitochondrial calcium overload were detected in the GCs of Amh-cre; Foxj2 tg/tg mice. Mcu (encoding a mitochondrial calcium uniporter) was observed to be upregulated in the GCs of the Amh-cre; Foxj2 tg/tg mice and was a direct target of FOXJ2. Moreover, Mcu knockdown restored mitochondrial calcium homeostasis and reduced the apoptosis in the GCs of the Amh-cre; Foxj2 tg/tg mice and in KGN cells transfected with FOXJ2-overexpression lentivirus.https://doi.org/10.1186/s13048-025-01651-0Follicle developmentGranulosa cellsApoptosisMitochondrial calcium overloadFOXJ2MCU |
| spellingShingle | Yunxia Zhang Qiqian Wu Furong Bai Yanqin Hu Bufang Xu Yujie Tang Jingwen Wu Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload Journal of Ovarian Research Follicle development Granulosa cells Apoptosis Mitochondrial calcium overload FOXJ2 MCU |
| title | Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload |
| title_full | Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload |
| title_fullStr | Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload |
| title_full_unstemmed | Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload |
| title_short | Granulosa cell-specific FOXJ2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload |
| title_sort | granulosa cell specific foxj2 overexpression induces premature ovarian insufficiency by triggering apoptosis via mitochondrial calcium overload |
| topic | Follicle development Granulosa cells Apoptosis Mitochondrial calcium overload FOXJ2 MCU |
| url | https://doi.org/10.1186/s13048-025-01651-0 |
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