State Key Laboratory of Swine and Poultry Breeding Industry, Key Laboratory of Livestock and Poultry Multi-omics, Ministry of Agriculture and Rural Affairs, and Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, China.
Animal Breeding and Genetics Key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, China.
Theriogenology. 2024 Dec;230:250-262. doi: 10.1016/j.theriogenology.2024.09.027. Epub 2024 Sep 26.
Normal follicular development is the basis for ovulation in poultry. Our previous sequencing analysis revealed a high expression of miR-24-3p in chicken follicles from degenerated ovaries, suggesting that miR-24-3p may modulate follicular development. Hence, this study investigated the specific mechanisms of miR-24-3p in regulating chicken follicular development. The results revealed that the proliferation, lipid synthesis, and progesterone secretion were significantly inhibited after miR-24-3p overexpression in chicken granulosa cells, vice versa by miR-24-3p knockdown. Dual-specificity phosphatase 16 (DUSP16) and thousand and one amino acid kinase 1 (TAOK1) were identified as potential target genes of miR-24-3p. Further validation revealed that knockdown of DUSP16 and TAOK1 suppressed proliferation, lipid synthesis, and progesterone secretion in chicken granulosa cells. Moreover, we observed that miR-24-3p, along with knockdown of DUSP16 and TAOK1, increased the phosphorylation levels of extracellular signal-regulated kinases 1 and 2 (ERK1/2). Our previous study proved that activation of ERK1/2 inhibited lipid synthesis and progesterone secretion of chicken granulosa cells. In summary, we demonstrated that miR-24-3p targeting DUSP16 and TAOK1 disrupts lipid synthesis and progesterone secretion via ERK1/2 signaling pathway in chicken granulosa cells in vitro. These results may provide a new theoretical basis for resolving miRNAs regulation on reproductive performance of chickens.
正常的卵泡发育是禽类排卵的基础。我们之前的测序分析表明，在退化卵巢的鸡卵泡中，miR-24-3p 表达水平较高，提示 miR-24-3p 可能调节卵泡发育。因此，本研究探讨了 miR-24-3p 调节鸡卵泡发育的具体机制。结果表明，在鸡颗粒细胞中过表达 miR-24-3p 后，增殖、脂质合成和孕激素分泌明显受到抑制，而 miR-24-3p 敲低则相反。双特异性磷酸酶 16（DUSP16）和千氨酸激酶 1（TAOK1）被鉴定为 miR-24-3p 的潜在靶基因。进一步验证表明，DUSP16 和 TAOK1 的敲低抑制了鸡颗粒细胞的增殖、脂质合成和孕激素分泌。此外，我们观察到 miR-24-3p 与 DUSP16 和 TAOK1 的敲低一起增加了细胞外信号调节激酶 1 和 2（ERK1/2）的磷酸化水平。我们之前的研究证明，ERK1/2 的激活抑制了鸡颗粒细胞的脂质合成和孕激素分泌。综上所述，我们证明了 miR-24-3p 通过靶向 DUSP16 和 TAOK1 破坏了鸡颗粒细胞中 ERK1/2 信号通路的脂质合成和孕激素分泌。这些结果可能为解决 miRNA 对鸡生殖性能的调节提供新的理论依据。
