Molecular mechanisms of avian neural crest cell migration on fibronectin and laminin.
Molecular mechanisms of avian neural crest cell migration on fibronectin and laminin.
复制标题
禽神经嵴细胞在纤连蛋白和层粘连蛋白上迁移的分子机制。
DOI：
10.1016/0012-1606(89)90144-9
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发表时间：
1989
影响因子：
2.7
通讯作者：
Bronner-Fraser,M
中科院分区：
文献类型：
--
作者：
Perris,R;Paulsson,M;Bronner-Fraser,M
我们已经研究了鸟类神经嵴细胞与纤维连接蛋白和层粘连蛋白的分子相互作用，在其最初的迁移过程中从神经管。一个105 kDa的纤连蛋白的蛋白水解片段，包括定义的细胞结合域（65 kDa）促进迁移的神经嵴细胞的完整分子的相同程度。神经嵴细胞迁移的完整的纤连蛋白和105-kDa的片段可逆地抑制含RGD肽。含有RGDS细胞附着位点的11.5-kDa片段也能够支持迁移，而对应于所定义的细胞结合结构域的相邻N-末端部分的50-kDa片段不利于神经嵴细胞运动。除了推定的“细胞结合域”，神经嵴细胞能够迁移的31-kDa片段对应的C-末端肝素结合（II）纤连蛋白的区域，并抑制其迁移外源性肝素，但不是RGDS肽。肝素增强了RGDS肽对完整纤连蛋白的抑制作用，但对105 kDa片段没有。纯化层粘连蛋白的基板上，鸟类神经嵴细胞迁移的程度是最大的，在相对较低的基板浓度和降低在较高的浓度。当糖蛋白与巢蛋白复合时，层粘连蛋白作为迁移底物的效率增强。此外，偶联层粘连蛋白巢蛋白复合物的IV型胶原蛋白或低密度硫酸乙酰肝素蛋白多糖进一步增加细胞分散，而单独的巢蛋白或蛋白多糖不能刺激迁移和IV型胶原蛋白是一个显着效率较低的迁移基板比层粘连蛋白巢蛋白。层粘连蛋白巢蛋白上的神经嵴细胞迁移不受RGDS或含YIGSR的肽的影响，但在加入肝素后减少了35%。层粘连蛋白的主要运动促进活性定位于E8结构域，具有与N末端E3结构域不同的肝素结合活性。添加肝素后，E8片段上的迁移减少了>70%。E1′片段支持最小程度的迁移，这是RGD敏感和肝素不敏感的，而主要的肝素结合E3片段和细胞粘附P1片段完全不允许细胞移动。将层粘连蛋白巢蛋白底物与抗E8片段的抗血清预孵育，而不与抗E1′或E4片段的抗血清预孵育，有效地减少了复合物上的迁移，进一步表明E8结构域是层粘连蛋白的主要运动促进区域。我们的结论是，最初的神经嵴细胞迁移纤连蛋白发生主要是通过与RGDS网站内的细胞结合域的相互作用，而其他潜在的附件/运动促进网站可能会起到稳定细胞-纤连蛋白的联系。层粘连蛋白上的神经嵴细胞迁移主要由E8结构域介导。该结构域的效率以及其他潜在的运动促进结构域刺激细胞运动的能力可能受到层粘连蛋白与其他细胞外基质分子结合的影响。
We have examined the molecular interactions of avian neural crest cells with fibronectin and lamininin vitroduring their initial migration from the neural tube. A 105-kDa proteolytic fragment of fibronectin encompassing the defined cell-binding domain (65 kDa) promoted migration of neural crest cells to the same extent as the intact molecule. Neural crest cell migration on both intact fibronectin and the 105-kDa fragment was reversibly inhibited by RGD-containing peptides. The 11.5-kDa fragment containing the RGDS cell attachment site was also able to support migration, whereas a 50-kDa fragment corresponding to the adjacent N-terminal portion of the defined cell-binding domain was unfavorable for neural crest cell movement. In addition to the putative “cell-binding domain,” neural crest cells were able to migrate on a 31-kDa fragment corresponding to the C-terminal heparin-binding (II) region of fibronectin, and were inhibited in their migration by exogenous heparin, but not by RGDS peptides. Heparin potentiated the inhibitory effect of RGDS peptides on intact fibronectin, but not on the 105-kDa fragment. On substrates of purified laminin, the extent of avian neural crest cell migration was maximal at relatively low substrate concentrations and was reduced at higher concentrations. The efficiency of laminin as a migratory substrate was enhanced when the glycoprotein occurred complexed with nidogen. Moreover, coupling of the laminin-nidogen complex to collagen type IV or the low density heparan sulfate proteoglycan further increased cell dispersion, whereas isolated nidogen or the proteoglycan alone were unable to stimulate migration and collagen type IV was a significantly less efficient migratory substrate than laminin-nidogen. Neural crest cell migration on laminin-nidogen was not affected by RGDS nor by YIGSR-containing peptides, but was reduced by 35% after addition of heparin. The predominant motility-promoting activity of laminin was localized to the E8 domain, possessing heparin-binding activity distinct from that of the N-terminal E3 domain. Migration on the E8 fragment was reduced by >70% after addition of heparin. The E1′ fragment supported a minimal degree of migration that was RGD-sensitive and heparin-insensitive, whereas the primary heparin-binding E3 fragment and the cell-adhesive P1 fragment were entirely nonpermissive for cell movement. Preincubation of laminin-nidogen substrates with antisera against the E8 fragment, but not against the E1′ or the E4 fragment, potently reduced migration on the complex, further suggesting that the E8 domain is the predominant motility-promoting region of laminin. We conclude that initial neural crest cell migration on fibronectin occurs primarily through an interaction with the RGDS site within the cell-binding domain, whereas other potential attachment/motility-promoting sites may act to stabilize cell-fibronectin linkages. Neural crest cell migration on laminin is primarily mediated by the E8 domain. The efficiency of this domain as well as the ability of other potential motility-promoting domains to stimulate cell movement may be influenced by the association of laminin with other extracellular matrix molecules.