Interaction of embryonic corneal epithelium with exogenous collagen, laminin, and fibronectin: role of endogenous protein synthesis.
Interaction of embryonic corneal epithelium with exogenous collagen, laminin, and fibronectin: role of endogenous protein synthesis.
复制标题
胚胎角膜上皮与外源胶原、层粘连蛋白和纤连蛋白的相互作用：内源蛋白质合成的作用。
DOI：
10.1016/0012-1606(82)90154-3
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发表时间：
1982
影响因子：
2.7
通讯作者：
Hay,ED
中科院分区：
文献类型：
--
作者：
Sugrue,SP;Hay,ED
以前，我们已经表明，胚胎角膜上皮细胞能够与外源性胶原蛋白，层粘连蛋白，和纤连蛋白的可溶性形式，其中每一个导致孤立的上皮细胞培养在微孔过滤器停止起泡，重组的基础细胞骨架，并扁平化。在这里，我们研究参与内源性衍生的细胞外基质（ECM）分子的基底上皮细胞表面与添加的ECM分子的相互作用。我们在这里证明，离体禽角膜上皮细胞培养的微孔过滤器是能够合成胶原和层粘连蛋白，但不是纤连蛋白。为了检查上皮是否能够直接与外源性ECM组分相互作用，或者是否需要产生接头分子，用放线菌酮（CHX）抑制上皮蛋白合成。然后，在CHX存在下的起泡上皮面对在过滤器下添加到培养基中的可溶性ECM分子;这样的上皮能够与胶原蛋白和层粘连蛋白相互作用，并响应于胶原蛋白和层粘连蛋白而变平。然而，这种受抑制的上皮细胞在纤连蛋白存在下继续流血的。我们接下来使用1-氮杂环丁烷-4-羧酸（LACA）干扰胶原蛋白分泌。暴露于LACA的上皮仍然能够与胶原蛋白和层粘连蛋白相互作用，但不能与纤连蛋白相互作用，表明依赖于胶原蛋白分泌。这些结果表明，纤连蛋白需要一个连接蛋白，可能是胶原蛋白，与基底上皮表面相互作用，而胶原蛋白和层粘连蛋白可以直接与细胞表面相互作用，将基底细胞骨架转化为典型的分化角膜上皮原位皮质垫。
Previously, we have shown that the embryonic corneal epithelium is capable of interacting with exogenous collagen, laminin, and fibronectin in soluble form, each of which causes isolated epithelium cultured on Millipore filter to stop blebbing, reorganize the basal cytoskeleton, and flatten. Here we examine the involvement of endogenously derived extracellular matrix (ECM) molecules in the interaction of the basal epithelial cell surface with the added ECM molecules. We demonstrate here that the isolated avian corneal epithelium cultured on Millipore filter is capable of synthesizing collagens and laminin, but not fibronectin. To examine whether the epithelium is capable of interacting directly with exogenous ECM components or if there is the necessity for production of a linker molecule, epithelial protein synthesis was inhibited with cycloheximide (CHX). The blebbing epithelium in the presence of CHX was then confronted with soluble ECM molecules added to the medium under the filter; such epithelia are able to interact with, and flatten in response to, both collagen and laminin. However, such inhibited epithelia continue to bled in the presence of fibronectin. We next usedl-azetidine-4-carboxylic acid (LACA) to interfere with collagen secretion. Epithelia exposed to LACA are still capable of interacting with collagen and laminin, but not fibronectin, indicating a dependence on collagen secretion. These results suggest that fibronectin requires a linker protein, probably collagen, to interact with the basal epithelial surface, whereas both collagen and laminin may interact directly with the cell surface to transform the basal cytoskeleton into the cortical mat typical of differentiating corneal epitheliumin situ.