Mochii M, Yoshida S, Morita K, Kohara Y, Ueno N
Department of Developmental Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan.
Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15020-5. doi: 10.1073/pnas.96.26.15020.
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.
转化生长因子-β家族成员在脊椎动物和无脊椎动物的身体模式形成中发挥着关键作用。一个与转化生长因子-β相关的基因dbl-1，已被证明在秀丽隐杆线虫中调节体长和雄性射线模式。我们通过差异杂交筛选了阵列cDNA，以鉴定DBL-1信号的下游靶基因。代表7584个独立基因的秀丽隐杆线虫cDNA被高密度排列在尼龙膜上，并与从野生型、dbl-1、sma-2和lon-2蠕虫的mRNA合成的(33)P标记的DNA探针杂交。对代表杂交DNA的所有斑点的信号进行定量，并在各菌株之间进行比较。筛选鉴定出22个和2个克隆，分别受DBL-1信号的正向和负向调控。Northern杂交证实了大多数克隆的表达谱，表明使用阵列cDNA进行差异杂交具有良好的可靠性。原位杂交分析揭示了每个克隆的时空表达模式，并表明至少有四个基因，包括DBL-1的I型受体基因sma-6，受DBL-1信号的转录调控。
