Shin Y et al. (2005), Identification of neural genes using Xenopus DN...

XB-ART-2554

Dev Dyn 2005 Feb 01;2322:432-44. doi: 10.1002/dvdy.20229.

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Identification of neural genes using Xenopus DNA microarrays.

Shin Y , Kitayama A , Koide T , Peiffer DA , Mochii M , Liao A , Ueno N , Cho KW .

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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.

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Species referenced: Xenopus laevis
Genes referenced: alcam ddit4 h2az1 h3c13 lrig3 nasp pdk1 pdk4 rgma sfrp2 slc27a3 sox11 vwc2l vwc2l.2 vwf

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	Figure 1. Expression patterns of novel neural genes. Whole-mount in situ hybridization was performed to confirm localization of transcripts in whole embryos. Gene expression patterns shown are at gastrula stage (stage 10.5), neurula stage (stage 18), tailbud stage (stage 30). Average fold induction from microarray experiments are presented as ldquo Fold. rdquo a, anterior; ba, branchial arches; c, cranial ganglia; cg, cement gland; d, dorsal ectoderm; nc, neural crest; np, neural plate; ov, otic vesicle; p, posterior; *, dorsal blastopore lip. Induction of these genes in animal cap ectoderm after inhibition of bone morphogenetic protein (BMP) signaling was confirmed by reverse transcriptase-polymerase chain reaction analysis.
	h3c13 (H3 clustered histone 13) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 18, dorsal view, anterior up.
	nasp (nuclear autoantigenic sperm protein (histone-binding)) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 30, lateral view, anterior left, dorsal up.
	pdk4 (pyruvate dehydrogenase kinase, isozyme 4) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 30, lateral view, anterior left, dorsal up.
	slc27a3 (solute carrier family 27 (fatty acid transporter), member 3) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 30, lateral view, anterior left, dorsal up.
	vwc2l.2 (von Willebrand factor C domain-containing protein 2-like, gene 2) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 30, lateral view, anterior left, dorsal up.