Systematic screening for signaling molecules expressed during somitogenesis by the signal sequence trap method

We describe a systematic screening to search for molecules that act as an extracellular signal during somitogenesis in vertebrates. Somitogenesis, which gives rise to segmented structures of axial bones and muscles, is a consequence of cooperative morphogenetic movements caused by precisely regulated cell and tissue interactions. We employed a strategy that combined subtractive hybridization to enrich paraxial mesoderm/somite-specific cDNAs and the signal sequence trap method, which selects signal sequence-containing molecules. Ninety-two independent cDNAs found to possess a putative signal sequence or a transmembrane domain are presented with a data base accession number for each. These clones include cDNAs which were previously identified with a function characterized, cDNAs previously identified with an undetermined function, and also cDNAs with no similarity to known sequences. Among them, 16 clones exhibited peculiar patterns of expression in the presomitic mesoderm/somites revealed by whole-mount and section in situ hybridization techniques, with some clones also being expressed in the forming neural tube. This is the first report in which an elaborate strategy combining three different screening steps was employed to identify signaling molecules relevant to a particular morphogenetic process.