Regulated expression of a novel laminin beta subunit during the development of the chick embryo

In order to define specific laminin variants implicated in organogenesis, we have undertaken a systematic search to detect and characterize novel laminin subunits, the expression of which is both developmentally regulated and tissue-specific. cDNA prepared from embryonic chick tissues was amplified by the polymerase chain reaction (PCR) with degenerate primers based on conserved sequences in domains V and VI of members of the laminin beta subunit family. Restriction mapping, cloning and sequencing of PCR products demonstrated a novel cDNA, the derived protein sequence of which displayed greatest homology with laminin beta subunits. However, the degree of amino acid divergence, comparison of sequence motifs and pattern of expression indicates that the cloned cDNA does not code for the avian orthologue of a previously characterized laminin beta subunit. Northern blot analysis showed that the expression of the 6-kb mRNA coding for the novel subunit was restricted to the skin. The mRNA was not detectable before day 5 of chick embryonic development, after which in situ hybridization showed expression only in surface ectoderm cells and subsequently in the epidermis. The developmentally regulated ectodermal expression of the novel beta subunit, prior to condensation of mesenchymal cells to form the dermis, is consistent with a specific role for this laminin isoform in the development and maintenance of the skin.