Damjanovski S et al. (1999), Spatial and temporal regulation of collagenases...

XB-ART-12635

Cell Res 1999 Jun 01;92:91-105. doi: 10.1038/sj.cr.7290009.

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Spatial and temporal regulation of collagenases-3, -4, and stromelysin -3 implicates distinct functions in apoptosis and tissue remodeling during frog metamorphosis.

Damjanovski S , Ishizuya-Oka A , Shi YB .

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Matrix metalloproteinases (MMPs) are a family of extracellular proteases capable of degrading various proteinaceous components of the extracellular matrix (ECM). They have been implicated to play important roles in a number of developmental and pathological processes, such as tumor metastasis and inflammation. Relatively few studies have been carried out to investigate the function of MMPs during postembryonic organ-development. Using Xenopus laevis development as a model system, we demonstrate here that three MMPs, stromelysin-3 (ST3), collagenases-3 (Col3), and Col4, have distinct spatial and temporal expression profiles during metamorphosis as the tadpole transforms into a frog. In situ hybridizations reveal a tight, but distinct, association of individual MMPs with tissue remodeling in the tail and intestine during metamorphosis. In particular, ST3 expression is strongly correlated with apoptosis in both organs as demonstrated by analyses of serial sections with in situ hybridization for ST3 mRNA and TUNEL (terminal deoxyribonucleotidyl transferase-mediated dUTP-biotin nick end labeling) for apoptosis, respectively. On the other hand, Col3 and Col4 are present in regions where extensive connective tissue remodeling take place. These results indicate that ST3 is likely to play a role in ECM-remodeling that facilitate apoptotic tissue remodeling or resorption while Col3 and Col4 appear to participate in connective tissue degradation during development.

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Species referenced: Xenopus laevis
Genes referenced: mmp1 mmp11 mmp13 tfcp2

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	Figure 2. Association of ST3 expression with larval epithelial cell death during intestinal remodeling. Stages 56 (A, B, C) and stage 60 (D, E, F) intestines were analyzed by in situ hybridization for ST3 expression (A, B, D, E) and by TUNEL assays for apoptotic cell detection (C, F). There was no detectable ST3 expression at stage 56 (B). ST3 expression was strong in all regions of the connective tissue at stage 60 but not in the epithelium or muscles (E). Likewise, at stage 56, there was no apoptotic signals (C), while there was many apoptotic signals, mainly within the epithelium, at stage 60 (arrows). Note that the control hybridization with the sense RNA probe did not detect any signal, as expected. ct, connective tissue; e, epithelium; l, lumen. White bars: 250 um.
	Figure 3. Collagenase-3, but not collagenase-4, can be detected by in situ hybridization during intestine metamorphosis. In situ hybridization for collagenase-3 (A, C) and collagenase-4 (B, D) at stage 56 (A, B) and stage 60 (C, D) demonstrated that collagenase-3 was expressed only at stage 60 and only in a subset of the connective tissue cells. ct, connective tissue; e, epithelium; l, lumen. White bars: 250 um
	Figure 5. Both collagenase-3 and collagenase-4 are abundantly expressed within the notochordal sheath during tail metamorphosis. In situ hybridization for collagenase-3 (A, C) and collagenase-4 (B, D) at stage 62 (A, B) and stage 64 (C, D) demonstrated that both mRNAs were detectable within the cells of the notochordal sheath through most of period of tail metamorphosis. Collagenase-3 was also detectable in the sub-epithelial fibroblasts at stage 62. n, notochord; ns, notochordal sheath; s, skin; sef, subepithelial fibroblasts. Bars in A, B, 100 mm; and in C, D: 200 um.