MMP-2 plays an essential role in producing epithelial-mesenchymal transformations in the avian embryo

To investigate the roles that matrix-degrading proteases may have in development of the chicken embryo, we documented the expression pattern of matrix metalloprotease-2 (MMP-2, 72-kDa type IV collagenase or gelatinase A) and perturbed its function in vitro and in vivo. MMP-2 is expressed as neural crest cells detach from the neural epithelium during an epithelial-mesenchymal transformation (EMT) but is rapidly extinguished as they disperse. It is also expressed in the sclerotome and in the dermis at the time that the EMT is initiated, and also as these cells migrate, and is down-regulated once motility has ceased. These patterns suggest that MMP-2 plays a role in cell motility during the EMT and during later morphogenesis. Inhibitors of MMPs, including BB-94 and TIMP-2 (tissue inhibitor of metalloprotease-2), prevent the EMT that generates neural crest cells, both in tissue culture and in vivo, but do not affect migration of the cells that have already detached from the neural tube. Similarly, knockdown of MMP-2 expression in the dorsal neural tube using antisense morpholino oligos perturbs the EMT, but also does not affect migration of neural crest cells after they have detached from the neural tube. On the other hand, when somites in culture are treated with TIMP-2, some mesenchymal cells are produced, suggesting that they undergo the EMT, but show greatly reduced migration through the collagen gel. MMP-2 is also expressed in mesenchyme where tissue remodeling is in progress, such as in the developing feather germs, in the head mesenchyme, in the lateral plate mesoderm, and in the limb dermis, especially in the regions where tendons are developing. Comparisons of these expression patterns in multiple embryonic tissues suggest a probable role for MMP-2 in the migration phase of the EMT, in addition to mesenchyme dispersion and tissue remodeling. (C) 2003 Wiley-Liss, Inc.