Normal skeletal pattern formation in chick limb bud with a mesenchymal hole is mediated by adjustment of cellular properties along the anterior?posterior axis in the limb bud
Normal skeletal pattern formation in chick limb bud with a mesenchymal hole is mediated by adjustment of cellular properties along the anterior?posterior axis in the limb bud
复制标题
具有间充质孔的鸡肢芽中正常骨骼模式的形成是通过沿着肢芽的前后轴调整细胞特性来介导的
DOI：
10.1016/j.ydbio.2021.12.016
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发表时间：
2022
影响因子：
2.7
通讯作者：
Wada Naoyuki
中科院分区：
文献类型：
--
作者：
Sato Yuki;Fujiwara Momoko;Nishino Haruka;Harada Rei;Kawasaki Eriko;Morimoto Ryo;Ohgo Shiro;Wada Naoyuki
鸡肢芽在发育的早期阶段，切除部分间充质团，形成一个洞，具有可塑性，可以重建正常的骨骼形态。为了了解洞闭合和重建正常肢体轴以重建正常肢体骨骼的细节，我们专注于洞修复和肢体修复过程中的细胞和分子变化。我们从鸡后肢芽（第23期）的内侧区域切除了一个立方体形的间充质细胞团，并观察了以下形态发生。切除后15 h，骨洞闭合，随后恢复肢芽形态，48 h时软骨模式基本恢复。间充质细胞的谱系分析表明，细胞在孔的前部和后部的边缘毗邻孔关闭网站，而细胞在近端和远端边缘没有。为了研究孔修复过程中的细胞极性，我们分析了高尔基体相对于细胞核的细胞内定位。我们发现，高尔基体倾向于指向孔之间的细胞在前，后边缘，但不是在细胞之间在相同的位置在正常肢芽或细胞在近端和远端孔边缘。与对照侧相比，移植侧肢体芽细胞增殖频率维持不变，Tbx 3表达由原来局限于肢体芽前后缘的表达向内侧暂时扩大，随着肢体重建的进行，Tbx 3阴性细胞重新出现在肢体芽内侧区域。因此，间充质孔闭合和肢体重建主要是由前孔边缘和后孔边缘的细胞介导的。这些结果表明，细胞特性的调整沿着前后轴是至关重要的恢复肢体损伤和重建正常的骨骼模式。
The chick limb bud has plasticity to reconstruct a normal skeletal pattern after a part of mesenchymal mass is excised to make a hole in its early stage of development. To understand the details of hole closure and re-establishment of normal limb axes to reconstruct a normal limb skeleton, we focused on cellular and molecular changes during hole repair and limb restoration. We excised a cube-shaped mass of mesenchymal cells from the medial region of chick hindlimb bud (stage 23) and observed the following morphogenesis. The hole had closed by 15 h after excision, followed by restoration of the limb bud morphology, and the cartilage pattern was largely restored by 48 h. Lineage analysis of the mesenchymal cells showed that cells at the anterior and posterior margins of the hole were adjoined at the hole closure site, whereas cells at the proximal and distal margins were not. To investigate cell polarity during hole repair, we analyzed intracellular positioning of the Golgi apparatus relative to the nuclei. We found that the Golgi apparatus tended to be directed toward the hole among cells at the anterior and posterior margins but not among cells at identical positions in normal limb buds or cells at the proximal and distal hole margins. In the manipulated limb buds, the frequency of cell proliferation was maintained compared with the control side.Tbx3expression, which was usually restricted to anterior and posterior margins of the limb bud, was temporarily expanded medially and then reverted to a normal pattern as limb reconstruction proceeded, withTbx3negative cells reappearing in the medial regions of the limb buds. Thus, mesenchymal hole closure and limb reconstruction are mainly mediated by cells at the anterior and posterior hole margins. These results suggest that adjustment of cellular properties along the anteroposterior axis is crucial to restore limb damage and reconstruct normal skeletal patterns.