Identification of transforming growth factor-beta- regulated genes in caenorhabditis elegans by differential hybridization of arrayed cDNAs.
Members of the transforming growth factor-beta family play critical roles in body patterning, in both vertebrates and invertebrates. One transforming growth factor-beta-related gene, dbl-1, has been shown to regulate body length and male ray patterning in Caenorhabditis elegans. We screened arrayed cDNAs to identify downstream target genes for the DBL-1 signaling by using differential hybridization. C. elegans cDNAs representing 7,584 independent genes were arrayed on a nylon membrane at high density and hybridized with (33)P-labeled DNA probes synthesized from the mRNAs of wild-type, dbl-1, sma-2, and lon-2 worms. Signals for all the spots representing hybridized DNA were quantified and compared among strains. The screening identified 22 and 2 clones, which were positively and negatively regulated, respectively, by the DBL-1 signal. Northern hybridization confirmed the expression profiles of most of the clones, indicating good reliability of the differential hybridization using arrayed cDNAs. In situ hybridization analysis revealed the spatial and temporal expression patterns of each clone and showed that at least four genes, including the gene for the type I receptor for DBL-1, sma-6, were transcriptionally regulated by the DBL-1 signal.
Mochii
M
M
Department of Developmental Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan. mochii@nibb.ac.jp
Yoshida
S
S
Morita
K
K
Kohara
Y
Y
Ueno
N
N
D000818:Animals; D015152:Blotting, Northern; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins; D018076:DNA, Complementary / Q000302:isolation & purification; D015870:Gene Expression; D020869:Gene Expression Profiling; D017238:Genes, Helminth; D017403:In Situ Hybridization; D007422:Intestines / Q000254:growth & development; D008297:Male; D009479:Neuropeptides / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D015398:Signal Transduction; D014018:Tissue Distribution; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D029742:Caenorhabditis elegans Proteins; D018076:DNA, Complementary; C117509:Dbl-1 protein, C elegans; D009479:Neuropeptides; D016212:Transforming Growth Factor beta