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Regulation of bone morphologenetic protein 4/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes
ZHANG Hai-yu ZHANG Xiao-li JI Shu-fang BING Lu-jun HAO Jing*
Acta Anatomica Sinica ›› 2014, Vol. 45 ›› Issue (3) : 375-382.
Regulation of bone morphologenetic protein 4/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes
Objective To explore the effect and mechanism of the bone morphologenetic protein 4 (BMP4)/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes. Methods Three-day-old Kunming mouse ovarine tissues were digested by the two-step enzymatic method to extract and purify oocytes. The cultured oocytes were divided into three groups: the normal culture medium (Con group), the medium with BMP4 (BMP4 group), and the medium with BMP4 and BMP4 inhibitor (BMP4+inhibitor group). TUNEL was used to examine the effects of BMP4 on the survival of the primordial follicle oocyte; Immunohistochemical staining and Real-time quantitative PCR were performed to investigate the expressions of p-Smad1/5/8, sohlh2, c-kit and foxo3a; siRNA interference, sohlh2 plasmid transfection and LY294002 treatments were performed to explore the mechanism of the BMP4/Smad signaling pathway on the apoptosis of oocytes. Results TUNEL results demonstrated that the ratio of apoptotic oocytes in BMP4 group was significantly lower than that in the Con group (P<0.05) and the BMP4+inhibitor group(P<0.05); BMP4 significantly promoted the nuclear translocation of Smad and inhibited the nuclear translocation of foxo3a, the mRNA and protein levels of sohlh2 and c-kit remarkably increased in BMP4 group. The effect of BMP4 on the oocyte survival was significantly repressed after sohlh2 siRNA transfection. Sohlh2 overexpression up-regulated the expression of p-foxo3a, and this activity was abolished by LY 294002. Conclusion BMP4/Smad signaling pathway may inhibit primordial follicle oocyte apoptosis, via up-regulation of the expression of sohlh2 and c-kit, and then down-regulation of the nuclear translocation of foxo3a.
Bone morphologenetic protein 4/Smad signaling pathway / Oocyte / Sohlh2 / TUNEL / Immunohistochemistry / Mouse
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