Regulation of bone morphologenetic protein 4/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes
10.3969/j.issn.0529-1356.2014.03.015
- VernacularTitle:骨形态发生蛋白4／Smad 信号通路在小鼠原始卵泡卵母细胞凋亡中的调控作用
- Author:
Haiyu ZHANG
;
Xiaoli ZHANG
;
Shufang JI
;
Lujun BING
;
Jing HAO
- Publication Type:Journal Article
- Keywords:
Bone morphologenetic protein 4/Smad signaling pathway;
Oocyte;
Sohlh2;
TUNEL;
Immunohistochemistry;
Mouse
- From:
Acta Anatomica Sinica
2014;(3):375-382
- CountryChina
- Language:Chinese
-
Abstract:
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 BMP 4 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 BMP 4/Smad signaling pathway on the apoptosis of oocytes . Results TUNEL results demonstrated that the ratio of apoptotic oocytes in BMP 4 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 foxo 3a, 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 sohlh 2 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 foxo 3a.
