Salvetti A, Rossi L, Deri P, Batistoni R
Dipartimento di Fisiologia e Biochimica, Università di Pisa, Pisa, Italy.
Dev Dyn. 2000 Aug;218(4):603-14. doi: 10.1002/1097-0177(2000)9999:9999<::AID-DVDY1016>3.0.CO;2-C.
The minichromosome maintenance (MCM2-7) gene family encodes conserved proteins, which are essential for DNA replication licensing in eukaryotes. They are abundant in proliferating cells, and specific MCM transcripts undergo cell cycle-dependent oscillations. Here we report the characterization of a planarian MCM2 homologue, DjMCM2, which represents the first molecular marker for detecting proliferating cells in planarians. DjMCM2-expressing cells are broadly distributed in the mesenchymal space of the body, with the exception of the cephalic region, and are preferentially accumulated in the peripheral area of the dorso-lateral mesenchyme, along the anteroposterior axis. During regeneration, no DjMCM2 transcripts are observed within the blastema, according to the current view that this structure is not a proliferation site in planarians. Spatio-temporal changes in DjMCM2 RNA expression pattern in the stump parallel blastema growth, coordinately with the orientation of the cut. X-ray irradiation results in the disappearance of DjMCM2 expression, thus confirming that these transcripts are detected specifically in proliferating cells, visualized as neoblasts by in situ hybridization in dissociated cells. In addition to neoblasts, rare large DjMCM2-expressing cells are observed in macerates of tissues excised just below the wound, suggesting that cell types other than neoblasts may be sporadically recruited for proliferation in planarians.
微小染色体维持蛋白(MCM2 - 7)基因家族编码保守蛋白,这些蛋白对于真核生物中的DNA复制起始是必不可少的。它们在增殖细胞中含量丰富,并且特定的MCM转录本会经历细胞周期依赖性振荡。在此,我们报道了涡虫MCM2同源物DjMCM2的特性,它是检测涡虫中增殖细胞的首个分子标记。表达DjMCM2的细胞广泛分布于身体的间充质空间,但头部区域除外,并且优先沿着前后轴聚集在背外侧间充质的周边区域。在再生过程中,根据目前认为胚基不是涡虫增殖位点的观点,在胚基内未观察到DjMCM2转录本。DjMCM2 RNA表达模式在残端的时空变化与胚基生长平行,并与切割方向协调一致。X射线照射导致DjMCM2表达消失,从而证实这些转录本在增殖细胞中被特异性检测到,在解离细胞中通过原位杂交可将其可视化为新生细胞。除了新生细胞外,在伤口下方刚切除的组织匀浆中还观察到罕见的、表达DjMCM2的大细胞,这表明在涡虫中,除新生细胞外的其他细胞类型可能偶尔被募集用于增殖。