Bevan S G, Southey M C, Armes J E, Venter D J, Newgreen D F
Embryology LAboratory, Murdoch Institute, Parkville, Victoria, Australia.
Genomics. 1996 Dec 1;38(2):206-14. doi: 10.1006/geno.1996.0617.
The neural crest is of fundamental importance in the developments of the head and peripheral nervous system, in the evolution of the vertebrates, and clinically because it gives rise to developmental abnormalities and neoplasms in humans. We have established a resource for studying the development of the neural crest by systematically constructing cDNA libraries from spatiotemporally exact neural crest and related cell populations. Neural crest populations were obtained from vagal and thoracic axial levels and from branchial arches, at premigratory and early and late migratory stages, at localization stages, and after differentiation into dorsal root ganglion cells, Schwann cells, sympathetic neurons, adrenal medullary cells, and melanocytes. Libraries were constructed using several methods developed to approach the issue of making representative libraries from small amounts of tissue. The fidelity and usefulness of the libraries were tested, and this revealed that they expressed a variety of sequences such as integrins, CAMs, growth factors and their receptors, protein-tyrosine kinases, and phosphatases. Differential display also revealed a unique combination of cDNA species. We then selected libraries spatiotemporally appropriate for epithelium-mesenchyme transformation and probed for TGF-beta-related sequences. As anticipated, we confirmed the presence of TGF-beta 2 and dorsalin-1 but could not detect TGF-beta 1. We also revealed new expression sites, defined by the origin of the libraries, of receptors known to be expressed elsewhere (Tsk 7l; TBRII). We anticipate that this collection of cDNA libraries will be of use in studying normal and abnormal neural crest development, by both homology searches and differential expression approaches, with spatiotemporal expression information being inherent in the initial screen.
神经嵴在头部和周围神经系统的发育、脊椎动物的进化以及临床上都具有至关重要的意义，因为它会导致人类出现发育异常和肿瘤。我们通过系统地从时空精确的神经嵴及相关细胞群体构建cDNA文库，建立了一个用于研究神经嵴发育的资源库。神经嵴群体取自迷走神经和胸段轴旁水平以及鳃弓，处于迁移前、迁移早期和晚期、定位阶段，以及分化为背根神经节细胞、施万细胞、交感神经元、肾上腺髓质细胞和黑素细胞之后。文库是使用几种为解决从小量组织构建代表性文库问题而开发的方法构建的。对文库的保真度和实用性进行了测试，结果表明它们表达了多种序列，如整合素、细胞黏附分子、生长因子及其受体、蛋白酪氨酸激酶和磷酸酶。差异显示还揭示了cDNA种类的独特组合。然后，我们选择了在时空上适合上皮-间充质转化的文库，并探测了与转化生长因子-β（TGF-β）相关的序列。正如预期的那样，我们证实了TGF-β2和背侧素-1的存在，但未检测到TGF-β1。我们还揭示了已知在其他地方表达的受体（Tsk 7l；TBRII）由文库来源定义的新表达位点。我们预计，通过同源性搜索和差异表达方法，利用初始筛选中固有的时空表达信息，这个cDNA文库集合将有助于研究正常和异常的神经嵴发育。
