COLLAGEN TYPE-VI IN NEURAL CREST DEVELOPMENT - DISTRIBUTION IN-SITU AND INTERACTION WITH CELLS IN-VITRO
COLLAGEN TYPE-VI IN NEURAL CREST DEVELOPMENT - DISTRIBUTION IN-SITU AND INTERACTION WITH CELLS IN-VITRO
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DOI：
10.1002/aja.1001980207
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发表时间：
1993-10-01
影响因子：
2.5
通讯作者：
BRONNERFRASER, M
中科院分区：
文献类型：
--
作者：
PERRIS, R;KUO, HJ;BRONNERFRASER, M
我们已经研究了VI型胶原蛋白（Col VI）在体内神经嵴发育过程中的时空分布及其在体外促进神经嵴细胞附着和迁移的能力。亲和纯化的抗血清和链特异性单克隆抗体对鸡胶原VI的免疫定位组织切片和免疫印迹。在最初的神经嵴细胞迁移的阶段，α 1（VI）和α 2（VI）链的免疫定位与基底膜的神经管，体节，脊索和外胚层，而没有免疫反应被视为α 3（VI）链。免疫印迹分析证实了α 1（VI）和α 2（VI）链的表达和缺乏可检测的免疫反应性的α 3（VI）链在这些早期阶段的神经嵴发育。相反，在先进的阶段的迁移和神经节形成后，表达的α 3（VI）链正好与α 1（VI）和α 2（VI）链在并置与基底膜，背根神经节周围，并在纤维状的安排在发展中的真皮和腹侧硬结。Col VI促进神经嵴细胞附着和迁移的能力在体外使用这些过程的定量测定进行了测试。原生微丝和孤立的四聚体的Col VI强烈促进神经嵴细胞的附着和迁移。神经嵴细胞粘附和迁移的最佳刺激依赖于Col VI的结构完整性，因为未折叠和拆解的α链仅微弱地促进细胞粘附，并且在支持细胞运动中几乎不活动。天然大分子组织的Col VI进一步的重要性进行了分析，在实验中，解离的四聚体被重新关联的Ca 2 +-和温度依赖性的自聚集。与原生微丝相反，这些低聚复合物在促进神经嵴细胞运动方面效果较差，但仍保留了刺激最大细胞附着的能力。结果表明，Col VI是细胞外基质沉积沿着神经嵴迁移途径的主要成分，在那里它可能参与细胞运动的调节，作为迁移底物。Col VI促进神经嵴细胞粘附和运动的能力高度依赖于天然大分子排列的有序性。(C)1993 Wiley-Liss，Inc.
We have examined the spatio-temporal distribution of collagen type VI (Col VI) during neural crest development in vivo and its ability to promote neural crest cell attachment and migration in vitro. An affinity purified antiserum and chain-specific monoclonal antibodies against chicken Col VI were employed to immunolocalize the collagen in tissue sections and by immunoblotting. At stages of initial neural crest cell migration, the al(VI) and alpha2(VI) chains were immunolocalized in apposition with basement membranes of the neural tube, somites, notochord and ectoderm, whereas no immunoreactivity was seen for the alpha3(VI) chain. Immunoblotting analysis confirmed the expression of alpha1(VI) and alpha2(VI) chains and the lack of detectable immunoreactivity for the alpha3(VI) chain at these early phases of neural crest development. Conversely, at advanced phases of migration and following gangliogenesis, expression of alpha3(VI) chain coincided with that of alpha1(VI) and alpha2(VI) chains in apposition with basement membranes, around the dorsal root ganglia, and in fibrillar arrangements within the developing dermis and ventral sclerotome. The ability of Col VI to promote neural crest cell attachment and migration was tested in vitro using quantitative assays for these processes. Both native microfilaments and isolated tetramers of Col VI strongly promoted neural crest cell attachment and migration. Optimal stimulation of neural crest cell adhesion and migration was dependent upon structural integrity of Col VI since unfolded and disassembled alpha chains only weakly promoted cell attachment and were virtually inactive in supporting cell movement. The importance of a native macromolecular organization of Col VI further was analyzed in experiments in which dissociated tetramers were reassociated by Ca2+- and temperature-dependent self-aggregation. In contrast to native microfilaments, these oligomeric complexes were less effective in promoting neural crest cell movement, but still retained the ability to stimulate maximal cell attachment. The results indicate that Col VI is a primary component of the extracellular matrix deposited along neural crest migratory pathways, where it may participate in the regulation of cell movement by functioning as a migratory substrate. The ability of Col VI to promote neural crest cell adhesion and motility is highly dependent upon maintainance of a native macromolecular arrangement. (C) 1993 Wiley-Liss, Inc.