Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, 117543 Singapore.
Curr Mol Med. 2012 Sep;12(8):1040-9. doi: 10.2174/156652412802480853.
Piwi controls the number of primordial germ cells (PGCs) via protecting maternal mRNA from decay and adult germ stem cell division in Drosophila. In mouse and zebrafish, piwi controls maintenance and differentiation of adult germ stem cell during gametogenesis. Whether piwi plays a role in PGC development of vertebrates remains unsolved. We addressed this issue by using medaka (Oryzias latipes) as a vertebrate model. Molecular cloning, sequence comparison and analyses of genomic organization and chromosome synteny led to the identification in this fish of a single piwi gene, called Opiwi. By RT-PCR analyses and in situ hybridization, the Opiwi transcript is maternally supplied and becomes restricted to PGCs and the central nervous system (CNS). Opiwi knockdown did not prevent PGC formation even in the absence of any somatic structures but did significantly reduce the number of PGCs in vivo and in vitro and affect the distribution of PGCs in developing embryos. Surprisingly, depletion of zygotic Opiwi severely and specifically affected PGC migration. We conclude that Opiwi is required not only for determining the PGC number but also for controlling PGC migration. Our results demonstrate that piwi plays a generally conserved role in germ cell development from Drosophila to vertebrate and a specific role in PGC migration.
Piwi 通过保护母体 mRNA 不降解和调控果蝇成体生殖干细胞分裂来控制原始生殖细胞(PGC)的数量。在小鼠和斑马鱼中,piwi 调控配子发生过程中成体生殖干细胞的维持和分化。piwi 是否在脊椎动物的 PGC 发育中起作用尚不清楚。我们通过使用鱼类(斑马鱼)作为脊椎动物模型来解决这个问题。通过分子克隆、序列比较以及基因组组织和染色体同线性分析,在这种鱼类中鉴定出一个称为 Opiwi 的单一 piwi 基因。通过 RT-PCR 分析和原位杂交,Opiwi 转录本是母源提供的,并局限于 PGC 和中枢神经系统(CNS)。Opiwi 敲低即使在没有任何体细胞结构的情况下也不能阻止 PGC 的形成,但确实显著减少了体内和体外的 PGC 数量,并影响了发育胚胎中 PGC 的分布。令人惊讶的是,合子 Opiwi 的耗竭严重且特异性地影响了 PGC 的迁移。我们得出结论,Opiwi 不仅对于确定 PGC 的数量是必需的,而且对于控制 PGC 的迁移也是必需的。我们的结果表明,piwi 在从果蝇到脊椎动物的生殖细胞发育中发挥着普遍保守的作用,并且在 PGC 迁移中发挥着特定的作用。