Regulation of bone morphologenetic protein 4/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes

doi:10.3969/j.issn.0529-1356.2014.03.015

Abstract

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 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.

Key words: Bone morphologenetic protein 4/Smad signaling pathway, Oocyte, Sohlh2, TUNEL, Immunohistochemistry, Mouse

ZHANG Hai-yu ZHANG Xiao-li JI Shu-fang BING Lu-jun HAO Jing*. Regulation of bone morphologenetic protein 4/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes[J]. AAS, 2014, 45(3): 375-382.

References

［1］Sui XX, Fu YC, Luo LL, et al. Forkhead box transcription factor foxo3a involves in oocyte apoptosis in neonatal rat［J］.China Journal of Modem Medicine， 2007，17(12) : 1438-1441.(in Chinese)
隋旭霞，傅玉才，罗丽莉，等.Foxo3a转录因子参与卵母细胞的凋亡［J］.中国现代医学杂志, 2007，17(12) : 1438-1441.
［2］Ding XY, Zhang XL, Hao J, et al. Effects of BMP4/SMAD signaling pathway on mouse primordial follicle growth and survival via up-regulation of Sohlh2 and c-kit［J］. Mol Reprod Dev, 2013, 80(1): 70-78. 
［3］Shimasaki S, Zachow RJ, Li D, et al. A functional bone morphogenetic protein system in the ovary［J］. Proc Natl Acad Sci USA, 1999, 96(13):7282-7287.
［4］Shimisaki S, Moore RK, Otsuka F,et al.The bone morphogenetic protein system in mammalian reproduction ［J］. Endocr Rev, 2004,25(1):72-101. 
［5］Tanwar PS, O’Shea T, McFarlane JR. In vivo evidence of role of bone morphogenetic protein-4 in the mouse ovary［J］. Ani Reprod Sci, 2008, 106(3-4):232-240. 
［6］Hao J, Li TG, Qi XX, et al. WNT/β-catenin pathway up-regulates Stat3 and converges on LIF to prevent differentiation of mouse embryonic stem cells［J］. Dev Biol, 2006, 290 (1): 81-91.［7］Fan XT, Hang YJ, Cai WQ, et al. Expression of BMP4 in CNS neurons of the developing rat［J］. Acta Anatomica Sinica, 2003, 34(6):568-572. (in Chinese)
范晓棠, 黄云剑, 蔡文琴, 等. 骨形成蛋白4在大鼠中枢神经系统发育过程中的表达［J］.解剖学报, 2003, 34(6):568-572.
［8］Nilsson EE, Skinner MK. Bone morphogenetic protein-4 acts as an ovarian follicle survival factor and promotes primordial follicle development［J］. Biol Reprod, 2003, 69(4):1265-1272.
［9］Su N, Li Y, Wang WJ, et al. Effect of Bone morphogenetic protein-4 on human primordial follicles in vitro［J］. Journal of Sun Yat-Sen Unversity（Medial Science）, 2008, 29(6):676-679. (in Chinese)
苏宁, 李予, 王文军, 等. 骨形态发生蛋白4对人始基卵泡的作用［J］. 中山大学学报（医学版）, 2008, 29(6):676-679.
［10］Suzuki H, Ahn HW, Rajkovic A. SOHLH1 and SOHLH2 coordinate spermatogonial differentiation［J］. Dev Biol, 2012, 361(2):301-312.
［11］Jagarlamudi K, Rajkovic A. Oogenesis: transcriptional regulators and mouse models［J］. Mol Cell Endocrinol,2012, 356(1-2):31-39. 
［12］Barrios F, Filipponi D, Dolci S, et al. SOHLH1 and SOHLH2 control Kit expression during postnatal male germ cell development［J］. J Cell Sci, 2012, 125(6):1455-1464.
［13］Choi Y, Yuan D, Rajkovic A. Germ cell-specific transcripti onal regulator sohlh2 is essential for early mouse folliculogenesis and oocyte-specific gene expression［J］. Biol Reprod, 2008, 79(6):1176-1182.
［14］Toyoda S, Miyazaki T, Miyazaki S, et al. Sohlh2 affects differrentiation of KIT positive oocytes and spermatogonia［J］.Dev Biol, 2009, 325(1):238-248.
［15］Jin X, Han CS, Yu FQ, et al. Anti-apoptotic action of stem cell factor on oocytes in primordial follicles and its signal transduction［J］. Mol Rep Dev, 2004,70(1):82-90.
［16］Liu H, Luo LL, Qian YS, et al.FOXO3a is involved in the apoptosis of naked oocytes and oocytes of primordial follicles from neonatal rat ovaries［J］.Bioche Biophys Res Commun,2009,381(4):722-727.
［17］Gilley J，Coffer PJ，Ham J.FOXO transcription factors directly activate bim gene expression and promote apoptosis in sympathetic neurons［J］.J Cell Biol，2003，162(4)：613-622.
［18］Dijkers PF，Medema RH，Lammers JW，et al.Expression of the pro-apoptofic Bcl-2 family member Bim is regulated by the forkhead transcrip-fion factor FKHR-LI［J］.Curt Biol, 2000，10(19)：1201-1204.
［19］Datta SR，Brunet A，Greenberg ME.Cellular survival：a play in three Akts［J］.Genes Dev，1999，13(22)：2905-2927.
［20］Burgering BM，Kops GJ.Cell cycle and death control：long live Forkheads［J］.Trends Biochem Sci，2002，27(7)：352-360.

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