Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan.
Dev Biol. 2010 May 15;341(2):429-43. doi: 10.1016/j.ydbio.2010.02.037. Epub 2010 Mar 15.
Planarian regeneration depends on the presence and precise regulation of pluripotent adult somatic stem cells named neoblasts, which differentiate to replace cells of any missing tissue. A characteristic feature of neoblasts is the presence of large perinuclear nonmembranous organelles named "chromatoid bodies", which are comparable to ribonucleoprotein structures found in germ cells of organisms across different phyla. In order to better understand regulation of gene expression in neoblasts, and potentially the function and composition of chromatoid bodies, we characterized homologues to known germ and soma ribonucleoprotein granule components from other organisms and analyzed their function during regeneration of the planarian Dugesia japonica. Expression in neoblasts was detected for 49 of 55 analyzed genes, highlighting the prevalence of post-transcriptional regulation in planarian stem cells. RNAi-mediated knockdown of two factors [ago-2 and bruli] lead to loss of neoblasts, and consequently loss of regeneration, corroborating with results previously reported for a bruli ortholog in the planarian Schmidtea mediterranea (Guo et al., 2006). Conversely, depletion mRNA turnover factors [edc-4 or upf-1], exoribonucleases [xrn-1 or xrn-2], or DEAD box RNA helicases [Djcbc-1 or vas-1] inhibited planarian regeneration, but did not reduce neoblast proliferation or abundance. We also found that depletion of cap-dependent translation initiation factors eIF-3A or eIF-2A interrupted cell cycle progression outside the M-phase of mitosis. Our results show that a set of post-transcriptional regulators is required to maintain the stem cell identity in neoblasts, while another facilitates proper differentiation. We propose that planarian neoblasts maintain pluripotency by employing mechanisms of post-transcriptional regulation exhibited in germ cells and early development of most metazoans.
涡虫再生依赖于多能成体体干细胞(称为成体神经干细胞)的存在和精确调控,这些干细胞可以分化为任何缺失组织的细胞。成体神经干细胞的一个特征是存在大的核周非膜细胞器,称为“染色质体”,与不同门的生物的生殖细胞中发现的核糖核蛋白结构相当。为了更好地理解成体神经干细胞中基因表达的调控,以及潜在的染色质体的功能和组成,我们鉴定了来自其他生物的已知生殖细胞和体细胞核糖核蛋白颗粒成分的同源物,并分析了它们在日本涡虫再生过程中的功能。在分析的 55 个基因中,有 49 个基因在成体神经干细胞中表达,这突出了转录后调控在涡虫干细胞中的普遍性。ago-2 和 bruli 两种因子的 RNAi 敲低导致成体神经干细胞的丢失,进而导致再生的丧失,这与先前在 Schmidtea mediterranea 涡虫中报道的 bruli 同源物的结果相吻合(Guo 等人,2006)。相反,mRNA 周转因子 [edc-4 或 upf-1]、外切核酸酶 [xrn-1 或 xrn-2] 或 DEAD 盒 RNA 解旋酶 [Djcbc-1 或 vas-1] 的耗竭抑制了涡虫再生,但不会减少成体神经干细胞的增殖或丰度。我们还发现,帽依赖性翻译起始因子 eIF-3A 或 eIF-2A 的耗竭会中断有丝分裂 M 期以外的细胞周期进程。我们的研究结果表明,一组转录后调控因子对于维持成体神经干细胞的干细胞特性是必需的,而另一种则促进适当的分化。我们提出,涡虫成体神经干细胞通过采用在生殖细胞和大多数后生动物的早期发育中表现出的转录后调控机制来维持多能性。
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