Skeletal regeneration in the brittle star Amphiura filiformis.
Skeletal regeneration in the brittle star Amphiura filiformis.
复制标题
脆性恒星Amphiura Filiformis中的骨骼再生。
DOI：
10.1186/s12983-016-0149-x
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发表时间：
2016
影响因子：
2.8
通讯作者：
Oliveri P
中科院分区：
文献类型：
--
作者：
Czarkwiani A;Ferrario C;Dylus DV;Sugni M;Oliveri P
海蛇尾在截肢后会再生整个手臂。由于转录组数据的可用性，丝状双足鱼现在可用于手臂再生的分子表征。先前的研究表明，已知参与棘皮动物骨骼形成的特定发育转录因子在丝状毛苋成体手臂再生过程中表达。然而，人们对骨骼形成的过程仍然知之甚少。在这里，我们展示了使用钙黄绿素染色、EdU 标记和原位杂交对再生过程中骨骼形态发生进行深入显微镜分析的结果。为了更好地比较不同的样本，我们基于活体动物中可识别的形态学标志并得到组织学分析的支持，提出了早期丝状线虫臂再生阶段的分期系统。我们发现，形成内骨骼的钙化针在再生过程中很早就出现在再生真皮层中。然后，这些骨针在再生后期成熟为分化手臂的复杂骨骼元件。真皮区域的间充质细胞表达骨骼标记基因Afi-c-lectin、Afi-p58b和Afi-p19；然而，EdU 标记显示这些真皮细胞不会增殖。尽管再生率存在差异，但丝状线虫臂使用远端化-嵌入模式通过一组一致的发育阶段进行再生。骨骼元素形成于不增殖的间充质细胞层中，因此必须由不同的来源提供。我们的工作为未来海蛇尾骨骼再生的细胞和分子研究奠定了基础。本文的在线版本 (doi:10.1186/s12983-016-0149-x) 包含补充材料，可供授权用户使用。
Brittle stars regenerate their whole arms post-amputation. Amphiura filiformis can now be used for molecular characterization of arm regeneration due to the availability of transcriptomic data. Previous work showed that specific developmental transcription factors known to take part in echinoderm skeletogenesis are expressed during adult arm regeneration in A. filiformis; however, the process of skeleton formation remained poorly understood. Here, we present the results of an in-depth microscopic analysis of skeletal morphogenesis during regeneration, using calcein staining, EdU labeling and in situ hybridization. To better compare different samples, we propose a staging system for the early A. filiformis arm regeneration stages based on morphological landmarks identifiable in living animals and supported by histological analysis. We show that the calcified spicules forming the endoskeleton first appear very early during regeneration in the dermal layer of regenerates. These spicules then mature into complex skeletal elements of the differentiated arm during late regeneration. The mesenchymal cells in the dermal area express the skeletal marker genes Afi-c-lectin, Afi-p58b and Afi-p19; however, EdU labeling shows that these dermal cells do not proliferate. A. filiformis arms regenerate through a consistent set of developmental stages using a distalization-intercalation mode, despite variability in regeneration rate. Skeletal elements form in a mesenchymal cell layer that does not proliferate and thus must be supplied from a different source. Our work provides the basis for future cellular and molecular studies of skeleton regeneration in brittle stars. The online version of this article (doi:10.1186/s12983-016-0149-x) contains supplementary material, which is available to authorized users.