Department of Biological Science, Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, Japan.
In Vitro Cell Dev Biol Anim. 2011 Aug;47(7):470-83. doi: 10.1007/s11626-011-9423-6. Epub 2011 May 26.
Larval-to-adult myogenic conversion occurs in the dorsal muscle but not in the tail muscle during Xenopus laevis metamorphosis. To know the mechanism for tail-specific suppression of adult myogenesis, response character was compared between adult myogenic cells (Ad-cells) and larval tail myogenic cells (La-cells) to a Sonic hedgehog (Shh) inhibitor, notochord (Nc) cells, and spinal cord (SC) cells in vitro. Cyclopamine, an Shh inhibitor, suppressed the differentiation of cultured Ad (but not La) cells, suggesting the significance of Shh signaling in promoting adult myogenesis. To test the possibility that Shh-producing axial elements (notochord and spinal cord) regulate adult myogenesis, Ad-cells or La-cells were co-cultured with Nc or SC cells. The results showed that differentiation of Ad-cells were strongly inhibited by Nc cells but promoted by SC cells. If Ad-cells were "separately" co-cultured with Nc cells without direct cell-cell interactions, adult differentiation was not inhibited but rather promoted, suggesting that Nc cells have two roles, one is a short-range suppression and another is a long-range promotion for adult myogenesis. Immunohistochemical analysis showed both notochord and spinal cord express the N-terminal Shh fragment throughout metamorphosis. The "spinal cord-promotion" and long-range effect by Nc cells on adult myogenesis is thus involved in Shh signaling, while the signaling concerning the short-range "Nc suppression" will be determined by future studies. Interestingly, these effects, "Nc suppression" and "SC promotion" were not observed for La-cells. Situation where the spinal cord/notochord cross-sectional ratio is quite larger in tadpole trunk than in the tail seems to contribute to trunk-specific promotion and tail-specific suppression of adult myogenesis during Xenopus metamorphosis.
在非洲爪蟾的变态过程中,成肌细胞向成体肌的转化发生在背肌,但不发生在尾肌。为了了解尾巴特异性抑制成体肌发生的机制,我们将成体肌细胞(Ad 细胞)和幼虫尾巴肌细胞(La 细胞)与 Sonic hedgehog(Shh)抑制剂、脊索(Nc)细胞和脊髓(SC)细胞在体外的反应特性进行了比较。Shh 抑制剂环巴胺抑制培养的 Ad(但不抑制 La)细胞的分化,表明 Shh 信号在促进成体肌发生中的重要性。为了验证产生 Shh 的轴向元件(脊索和脊髓)调节成体肌发生的可能性,我们将 Ad 细胞或 La 细胞与 Nc 细胞或 SC 细胞共培养。结果表明,Nc 细胞强烈抑制 Ad 细胞的分化,但促进 SC 细胞的分化。如果 Ad 细胞与 Nc 细胞“分开”共培养而没有直接的细胞-细胞相互作用,成体分化不会受到抑制,而是受到促进,这表明 Nc 细胞有两个作用,一个是短程抑制,另一个是长程促进成体肌发生。免疫组织化学分析显示,脊索和脊髓在整个变态过程中都表达 Shh 片段的 N 端。因此,Nc 细胞对成体肌发生的“脊髓促进”和长程作用涉及 Shh 信号,而关于短程“Nc 抑制”的信号将由未来的研究来确定。有趣的是,这些效应,“Nc 抑制”和“SC 促进”,在 La 细胞中没有观察到。在非洲爪蟾的变态过程中,脊髓/脊索横截面积比在尾部大得多的情况,似乎有助于成体肌在躯干特异性促进和尾部特异性抑制中的发生。