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Smad4 mediates Bmf involvement in sheep granulosa cell apoptosis
Mingna Li1, Weiwei Liang2, Caiye Zhu3
1Gansu Key Laboratory of Herbivorous Animal Biotechnology, Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China..
Abstract:
Bcl-2-modifying factor (Bmf) functions to mediate follicular atresia and oocyte growth in mice. It has been proven that TGF-β can induce Bmf expression via the Smad4 pathway in a variety of cells, and then induce cell apoptosis. Based on this, we hypothesized that Smad4 and Bmf may play important roles in the apoptosis of granulosa cells (GCs) in domestic animals. This study used small-tailed Han sheep follicular GCs cultured in vitro as a model system, and overexpression or interference experiments, to explore the biological roles of Bmf and reveal the preliminary regulatory mechanisms between Smad4 and Bmf in the process of GCs' apoptosis. We found that the proliferation rate of sheep GCs was significantly increased after the knockdown of Bmf, whereas overexpressing Bmf increased the apoptosis rate of GCs, results also verified by the expression patterns of PCNA, Bcl-2, and Bax genes. After the Smad4 knockdown, the apoptosis rate of GCs was increased, while the mRNA and protein expression of Bmf was significantly up-regulated. A rescue experiment verified that the Bmf knockdown could alleviate GCs' apoptosis induced by Smad4 knockdown. In conclusion, our study not only elucidated an important role for Bmf in the apoptosis of sheep GCs but also revealed a new regulatory pathway between Smad4 and Bmf in this process.
Insights
Bcl-2-modifying factor (Bmf) influences sheep granulosa cell (GC) apoptosis. Smad4 knockdown increases GC apoptosis and Bmf expression, while Bmf knockdown reduces apoptosis, revealing a novel Smad4-Bmf regulatory pathway.
Area of Science:
- Reproductive Biology
- Cell Biology
- Molecular Biology
Background:
- Bcl-2-modifying factor (Bmf) is involved in follicular atresia and oocyte growth in mice.
- Transforming growth factor-beta (TGF-β) can induce Bmf expression via the Smad4 pathway, leading to apoptosis.
- The roles of Smad4 and Bmf in domestic animal granulosa cell (GC) apoptosis are not well understood.
Purpose of the Study:
- To investigate the biological roles of Bmf in sheep GC apoptosis.
- To explore the regulatory mechanisms between Smad4 and Bmf in GC apoptosis.
Main Methods:
- Primary sheep GCs were cultured in vitro.
- Gene knockdown and overexpression techniques were employed for Bmf and Smad4.
- Cell proliferation and apoptosis rates were assessed.
- Gene and protein expression levels (PCNA, Bcl-2, Bax) were analyzed.
Main Results:
- Bmf knockdown increased sheep GC proliferation, while Bmf overexpression enhanced GC apoptosis.
- Smad4 knockdown led to increased GC apoptosis and significantly upregulated Bmf mRNA and protein expression.
- Bmf knockdown mitigated the apoptosis induced by Smad4 knockdown.
Conclusions:
- Bmf plays a significant role in regulating apoptosis of sheep granulosa cells.
- A novel regulatory pathway involving Smad4 and Bmf in GC apoptosis was identified.
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