EXPRESSION OF N-CADHERIN AND ALKALINE-PHOSPHATASE IN CHICK LIMB BUD MESENCHYMAL CELLS - REGULATION BY 1,25-DIHYDROXYVITAMIN D-3 OR TGF-BETA-1

When mesenchymal cells from the early chick limb bud (stage 23-24) are plated at high cell density they spontaneously undergo chondrogenesis implicating extensive cell-to-cell interactions. In the past it has b een shown that TGF-beta and vitamin D can influence this process and c an stimulate chondrogenesis. Given the importance of cell adhesion mol ecules during cellular interactions we decided to examine the effects of 1,25-dihydroxy-vitamin D-3 or TGF-beta on the expression of molecul es involved in cell-to-cell adhesion (N-cadherin) or cell-substrate ad hesion (alkaline phosphatase). Immunofluorescence demonstrated that N- cadherin was expressed in the mesenchymal cells and in the very early cartilage nodules but it was down-regulated in the older nodules. As s hown by Western blotting, the expression of N-cadherin declined as cho ndrogenesis proceeded and was affected in cultures treated with 1,25-d ihydroxyvitamin D-3 and TGB-beta 1. Alkaline phosphatase was also expr essed in the mesenchymal cells; these cells preferentially use an alte rnative transcript compared to the cartilage cells of the sternum. Thu s, our data suggest that the involvement of 1,25-dihydroxyvitamin D-3 in chondrogenesis could be mediated via regulation of cell adhesion. ( C) 1994 Academic Press, Inc.