Shin Yongchol, Kitayama Atsushi, Koide Tetsuya, Peiffer Daniel A, Mochii Makoto, Liao Arnold, Ueno Naoto, Cho Ken W Y
Department of Developmental and Cell Biology, University of California, Irvine, California, USA.
Dev Dyn. 2005 Feb;232(2):432-44. doi: 10.1002/dvdy.20229.
To isolate novel genes regulating neural induction, we used a DNA microarray approach. As neural induction is thought to occur by means of the inhibition of bone morphogenetic protein (BMP) signaling, BMP signaling was inhibited in ectodermal cells by overexpression of a dominant-negative receptor. RNAs were isolated from control animal cap explants and from dominant-negative BMP receptor expressing animal caps and subjected to a microarray experiment using newly generated high-density Xenopus DNA microarray chips representing over 17,000 unigenes. We have identified 77 genes that are induced in animal caps after inhibition of BMP signaling, and all of these genes were subjected to whole-mount in situ hybridization analysis. Thirty-two genes showed specific expression in neural tissues. Of the 32, 14 genes have never been linked to neural induction. Two genes that are highly induced by BMP inhibition are inhibitors of Wnt signaling, suggesting that a key step in neural induction is to produce Wnt antagonists to promote anterior neural plate development. Our current analysis also proves that a microarray approach is useful in identifying novel candidate factors involved in neural induction and patterning.
为了分离调控神经诱导的新基因，我们采用了DNA微阵列方法。由于认为神经诱导是通过抑制骨形态发生蛋白（BMP）信号传导来发生的，因此通过过表达显性负性受体在表皮细胞中抑制BMP信号传导。从对照动物帽外植体和表达显性负性BMP受体的动物帽中分离RNA，并使用代表超过17,000个单基因的新生成的高密度非洲爪蟾DNA微阵列芯片进行微阵列实验。我们已经鉴定出77个在抑制BMP信号传导后在动物帽中被诱导的基因，并且所有这些基因都进行了全胚胎原位杂交分析。32个基因在神经组织中显示出特异性表达。在这32个基因中，有14个基因从未与神经诱导相关联。两个被BMP抑制高度诱导的基因是Wnt信号传导的抑制剂，这表明神经诱导的一个关键步骤是产生Wnt拮抗剂以促进前神经板发育。我们目前的分析还证明，微阵列方法在鉴定参与神经诱导和模式形成的新候选因子方面是有用的。
