Perris R, Krotoski D, Bronner-Fraser M
Developmental Biology Center, University of California, Irvine 92717.
Development. 1991 Nov;113(3):969-84. doi: 10.1242/dev.113.3.969.
This study examines the spatiotemporal distribution of collagen (Col) types I-V and IX during neural crest development in vivo and their ability to support neural crest cell movement in vitro. Col I, III and IV were widespread throughout the embryo, including the neural crest migratory pathways, whereas Col II, V and IX preferentially localized to regions from which migrating neural crest cells were absent. Col I-IV and IX occurred both in association with basement membranes and within interstitial matrices, whereas Col V only was detected in juxtaposition to basement membranes. Although initially distributed throughout the rostrocaudal extent of the somitic sclerotome, Col I and III rearranged to the caudal portion with progressive neural crest cell migration through the rostral portion of the sclerotome. This rearrangement does not occur in neural crest-ablated embryos, suggesting that it is a direct consequence of neural crest cell migration. The perinotochordal matrix, avoided by neural crest cells, contained a metameric Col II/IX immunoreactivity along the rostrocaudal axis which alternated with that of Col I and III. In contrast, Col IV and V were not observed in this matrix, but lined the basement membranes of the notochord and ventrolateral neural tube. To determine their functional significance for neural crest cell migration in vivo, purified collagens were tested for their ability to promote neural crest cell motility in vitro. Neural crest cell migration on isolated collagens was most pronounced on Col I and IV, whereas Col II, V and the triple-helical fragment of Col VII were unable to support cell motility. Substrata created by copolymerization of Col I and fibronectin, or Col I and laminin-nidogen, supported cell motility better than Col I alone, whereas both Col V and a cartilage-type chondroitin sulfate proteoglycan reduced cell movement on Col I. Fibronectin bound to pre-immobilized monomeric Col I, II or V had a reduced ability to support neural crest cell movement when compared to fibronectin alone. A similar reduction was seen for Col IV bound to the low density heparan sulfate proteoglycan from the EHS mouse tumor. The results demonstrate that Col I-IX are differentially distributed in the early avian embryo. During neural crest development several of these collagens undergo dynamic reorganizations that correlate with the migration of neural crest cells. Furthermore, various collagens possess distinct abilities to support neural crest cell migration in vitro, and their migration-promoting activity can be modulated by their conformation and/or association with other matrix components.
本研究考察了体内神经嵴发育过程中I-V型和IX型胶原蛋白（Col）的时空分布及其在体外支持神经嵴细胞迁移的能力。I型、III型和IV型胶原蛋白广泛分布于整个胚胎，包括神经嵴迁移途径，而II型、V型和IX型胶原蛋白则优先定位于不存在迁移神经嵴细胞的区域。I-IV型和IX型胶原蛋白既与基底膜相关联，也存在于间质基质中，而V型胶原蛋白仅在与基底膜相邻处被检测到。尽管I型和III型胶原蛋白最初分布于体节生骨节的整个头尾范围，但随着神经嵴细胞逐渐通过生骨节的头侧部分迁移，它们会重新排列至尾侧部分。这种重新排列在神经嵴消融的胚胎中不会发生，这表明它是神经嵴细胞迁移的直接结果。神经嵴细胞避开的脊索周围基质沿头尾轴含有节段性的II型/IX型胶原蛋白免疫反应性，与I型和III型胶原蛋白的免疫反应性交替出现。相比之下，在该基质中未观察到IV型和V型胶原蛋白，但它们排列在脊索和腹外侧神经管的基底膜上。为了确定它们在体内对神经嵴细胞迁移的功能意义，对纯化的胶原蛋白进行了体外促进神经嵴细胞运动能力的测试。神经嵴细胞在分离的胶原蛋白上的迁移在I型和IV型胶原蛋白上最为明显，而II型、V型胶原蛋白和VII型胶原蛋白的三螺旋片段则无法支持细胞运动。由I型胶原蛋白与纤连蛋白或I型胶原蛋白与层粘连蛋白-巢蛋白共聚形成的基质比单独的I型胶原蛋白更能支持细胞运动，而V型胶原蛋白和软骨型硫酸软骨素蛋白聚糖都会减少神经嵴细胞在I型胶原蛋白上的运动。与单独的纤连蛋白相比，与预先固定的单体I型、II型或V型胶原蛋白结合的纤连蛋白支持神经嵴细胞运动的能力降低。与来自EHS小鼠肿瘤的低密度硫酸乙酰肝素蛋白聚糖结合的IV型胶原蛋白也出现了类似的降低。结果表明，I-IX型胶原蛋白在早期鸡胚中分布存在差异。在神经嵴发育过程中，其中几种胶原蛋白会发生动态重组，这与神经嵴细胞的迁移相关。此外，各种胶原蛋白在体外支持神经嵴细胞迁移的能力各不相同，它们的迁移促进活性可通过其构象和/或与其他基质成分的结合而受到调节。
