Rapid changes in the extracellular matrix accompany in vitro palatal shelf remodelling.
Rapid changes in the extracellular matrix accompany in vitro palatal shelf remodelling.
复制标题
细胞外基质的快速变化伴随着体外腭架重塑。
DOI：
10.1007/bf00191453
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Brinkley,L
中科院分区：
文献类型：
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作者：
Morris-Wiman,J;Brinkley,L
使用为早期胚胎培养开发的标准滚管培养方法，在体外模型中检查小鼠次级腭架抬高过程中的事件顺序和细胞外基质（ECM）成分的分布。在该培养系统中，妊娠第 13 天小鼠胎儿腭架重塑和重新定向相关的形态变化与体内观察到的相似。然而，在体内重新定向前24-30小时移植的标本中，移植后重塑迅速开始，并在4小时内完成显着的重新定向。直到体外18小时后，随着架子的生长，架子之间才发生中线接触。因此，在该体外模型中，与腭架重塑和重新定向相关的事件可以与与腭架生长相关的事件区分开来。我们使用该体外模型来表征与架重塑事件同时发生的 ECM 分布和积累的瞬时变化。我们的结果表明，在体外快速重塑过程中，通过免疫荧光染色观察到，胶原蛋白 III、纤连蛋白和透明质酸盐的相对分布在间充质区室的特定区域内减少。相比之下，间充质内I型胶原蛋白的分布增加，而生腱蛋白的分布没有显着变化。除生腱蛋白外，所有检查的分子都显示出基底膜内分布的变化。这些分布模式与体内更渐进的重塑过程中观察到的分布模式相似。在体外重塑过程中，ECM 的 [3H]-葡萄糖胺和 [3H]-脯氨酸标记成分的沉积（通过放射自显影观察）在培养的前 3 小时内沉积量最大。在此期间，标记的 ECM 在间充质和腭上皮基底膜的特定区域内积累。在随后的培养 3 小时内，摄取急剧减少，并且主要局限于腭上皮及其下面的基底膜。体外系统可以表征导致重新定向的架子重塑的早期事件。结果表明，重塑伴随着 ECM 在腭架特定区域的快速局部积累，以前认为在此过程中发挥了作用。
The sequence of events and the distribution of extracellular matrix (ECM) components was examined during mouse secondary palatal shelf elevation in an in vitro model using standard roller tube culture methods developed for the culture of early embryos. In this culture system, the morphological changes associated with remodelling and reorientation of the palatal shelves of gestational day 13 mouse fetuses were similar to those observed in vivo. However, in specimens explanted 24–30 h prior to reorientation in vivo, remodelling began rapidly after explantation, and significant reorientation was accomplished within 4 h. Midline contact between the shelves did not occur until after 18 h in vitro, concomitant with shelf growth. Therefore, in this in vitro model, events related to palatal shelf remodelling and reorientation can be distinguished from those related to shelf growth. We used this in vitro model to characterize the transient changes in ECM distribution and accumulation that occur concomitant with events in shelf remodelling. Our results show that, during rapid remodelling in vitro, the relative distributions of collagen III, fibronectin and hyaluronate, as visualized by immunofluorescent staining, decreased within specific regions of the mesenchymal compartment. In contrast, the distribution of collagen I within the mesenchyme increased, and the distribution of tenascin did not change significantly. All molecules examined, except tenascin, showed changes in distribution within the basement membrane. These patterns of distribution are similar to those observed during more gradual remodelling in vivo. During remodelling in vitro, the deposition of [3H]-glucosamine- and [3H]-proline-labelled components of the ECM, as visualized by autoradiography, was greatest during the first 3 h of culture. During this period, labelled ECM accumulated within specific regions of the mesenchyme and palatal epithelial basement membrane. Uptake was reduced dramatically during the subsequent 3 h in culture and was restricted mainly to the palatal epithelium and its underlying basement membrane. The in vitro system permitted the characterization of early events in shelf remodelling leading to reorientation. Results suggest that remodelling is accompanied by rapid, local accumulation of ECM in specific regions of the palatal shelf previously thought to play a role in the process.