Identification of neural genes using Xenopus DNA microarrays.
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.
Shin
Yongchol
Y
Department of Developmental and Cell Biology, University of California, Irvine, California, USA.
Kitayama
Atsushi
A
Koide
Tetsuya
T
Peiffer
Daniel A
DA
Mochii
Makoto
M
Liao
Arnold
A
Ueno
Naoto
N
Cho
Ken W Y
KW
D000818:Animals; D019521:Body Patterning; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis*; D002843:Chromatin / Q000378:metabolism; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D005799:Genes, Dominant; D017403:In Situ Hybridization; D009432:Neural Crest / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013997:Time Factors; D014982:Xenopus laevis
D019485:Bone Morphogenetic Proteins; D002843:Chromatin; D012313:RNA