Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
Gen Comp Endocrinol. 2012 Apr 1;176(2):173-81. doi: 10.1016/j.ygcen.2012.01.012. Epub 2012 Jan 28.
Basic fibroblast growth factor (bFGF or FGF2) plays diverse roles in regulating cell proliferation, migration and differentiation during embryo development. In this study, the effect of bFGF on ovarian germ cell development was investigated in the embryonic chicken by in vitro and in vivo experiments. Results showed that a remarkable decrease in bFGF expression in the ovarian cortex was manifested during meiosis progression. With ovary organ culture, we revealed that meiosis was initiated after retinoic acid (RA) treatment alone but was decreased after combined bFGF treatment that was detected by real time RT-PCR, fluorescence immunohistochemistry and Giemsa staining. Further, no significant difference in mRNA expression of either RA metabolism-related enzymes (Raldh2 and Cyp26b1) or RA receptors was displayed after bFGF challenge. This result suggests that the suppression of bFGF on meiosis was unlikely through inhibition of RA signaling. In addition, as a mitogen, bFGF administration increased germ cell proliferation (via BrdU incorporation) in cultured organ or cells in vitro and also in developing embryos in vivo. In contrast, blockade of bFGF action by SU5402 (an FGFR1 antagonist) or inhibition of protein kinase C signaling showed inhibited effect of bFGF on mitosis. In conclusion, bFGF suppresses RA-induced entry of germ cells into meiosis to ensure embryonic ovarian germ cells to maintain at undifferentiated status and accelerate germ cell proliferation by binding with FGFR1 involving PKC activation in the chicken.
碱性成纤维细胞生长因子(bFGF 或 FGF2)在胚胎发育过程中对细胞增殖、迁移和分化起着多样化的调节作用。本研究通过体外和体内实验,研究了 bFGF 对鸡胚胎卵巢生殖细胞发育的影响。结果表明,在减数分裂过程中,卵巢皮质中 bFGF 的表达明显下降。通过卵巢器官培养,我们发现单独用视黄酸(RA)处理即可启动减数分裂,但用 bFGF 联合处理后,减数分裂减少,实时 RT-PCR、荧光免疫组织化学和吉姆萨染色检测均显示这一结果。此外,bFGF 处理后,RA 代谢相关酶(Raldh2 和 Cyp26b1)或 RA 受体的 mRNA 表达没有明显差异。这一结果表明,bFGF 对减数分裂的抑制作用不太可能通过抑制 RA 信号通路。此外,作为有丝分裂原,bFGF 给药可增加体外培养器官或细胞中的生殖细胞增殖(通过 BrdU 掺入),也可增加体内发育胚胎中的生殖细胞增殖。相比之下,用 SU5402(FGFR1 拮抗剂)阻断 bFGF 作用或抑制蛋白激酶 C 信号转导,显示 bFGF 对有丝分裂的抑制作用。总之,bFGF 抑制 RA 诱导的生殖细胞进入减数分裂,以确保胚胎期卵巢生殖细胞保持未分化状态,并通过与 FGFR1 结合激活蛋白激酶 C 来加速生殖细胞增殖。