Tucker R P, McKay S E
Department of Neurobiology and Anatomy, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC 27103.
Development. 1991 Aug;112(4):1031-9. doi: 10.1242/dev.112.4.1031.
The extracellular matrix glycoprotein tenascin is concentrated in both the embryo and adult in regions where cell motility is taking place. For example, during avian neural crest morphogenesis tenascin is concentrated in the rostral half of the sclerotome, precisely where the neural crest cells themselves are found. Previous in vitro studies indicated that somite cells were the source of this tenascin, implying a role for tenascin in directing the ventral migration of neural crest cells and thus the establishment of the periodic arrangement of the PNS. In this study, we have used a cDNA probe to identify the source of tenascin found along the pathways of the neural crest using in situ hybridization. In tissue sections, individual cells found along the neural crest migratory pathways, both before entering the somites and within the somites, are strongly labelled by the tenascin cDNA. In vitro neural crest cells are more strongly labelled with the tenascin probe than somite cells. Finally, western blotting has been used to identify tenascin in culture medium conditioned by neural crest cells. This indicates that neural crest cells themselves are the source of much of the tenascin found lining their migratory pathways, and that interactions with somite cells may not be needed to induce the expression of tenascin. We have also studied the distribution of tenascin mRNA in the developing spinal cord and spinal ganglia. At embryonic days 7 and 10, tenascin cDNA hybridizes within cells that appear to be migrating from the ependymal layer to the white matter, as well as within cells in the dorsal roots.(ABSTRACT TRUNCATED AT 250 WORDS)
细胞外基质糖蛋白腱生蛋白集中于胚胎和成年个体中发生细胞迁移的区域。例如,在鸟类神经嵴形态发生过程中,腱生蛋白集中于体节的前半部分,正是神经嵴细胞所在的位置。先前的体外研究表明体节细胞是这种腱生蛋白的来源,这意味着腱生蛋白在引导神经嵴细胞向腹侧迁移从而建立周围神经系统的周期性排列中发挥作用。在本研究中,我们使用cDNA探针通过原位杂交来确定沿神经嵴路径发现的腱生蛋白的来源。在组织切片中,沿神经嵴迁移路径发现的单个细胞,无论是在进入体节之前还是在体节内部,都被腱生蛋白cDNA强烈标记。体外培养的神经嵴细胞比体节细胞被腱生蛋白探针标记得更强。最后,蛋白质印迹法已被用于鉴定神经嵴细胞条件培养基中的腱生蛋白。这表明神经嵴细胞自身是其迁移路径沿线发现的大部分腱生蛋白的来源,并且诱导腱生蛋白表达可能不需要与体节细胞相互作用。我们还研究了腱生蛋白mRNA在发育中的脊髓和脊神经节中的分布。在胚胎第7天和第10天,腱生蛋白cDNA在似乎从室管膜层迁移到白质的细胞内以及背根中的细胞内杂交。(摘要截短于250词)