title: A novel in-situ-zymography technique localizes gelatinolytic activity in human skin to mast cells.
abstract: Matrix-metallo-proteinases play a key role in cutaneous tissue remodeling and wound healing, and have been implicated as the rate-limiting factor in cutaneous tumor invasion and metastasis. We here describe a novel in-situ-zymographic method, which allows to directly localize sites of gelatinolytic activity in human skin. Gelatinolysis was detected through protein-hydrolysis in a 200 microm thick polyacrylamide gel underlying tissue sections. The lysis was substrate-dependent, demonstrated time- and temperature-dependent kinetics, and was inhibited by both EDTA and 1,10-phenanthroline. Normal and diseased skin sections demonstrated multiple focal points of gelatinolysis which co-localized with individual cells. Histochemically, these were shown to represent most likely mast cells (via AS-d-chloroacetate esterase staining and metachromasia). However, immunohistochemical staining for gelatinases A and B showed no immunoreactivity patterns that corresponded to the identified foci of gelatinolysis. The reported in-situ-zymographic technique offers a decisive advantage over immunohistochemistry, since it detects only the activated and catabolically relevant proteases, and provides further evidence for a role of mast cells in extracellular matrix remodeling.
authors.affiliation: Department of Dermatology, University of Erlangen, Germany.
authors.forename: N C
authors.initials: NC
authors.lastname: Krejci-Papa
authors.forename: R
authors.initials: R
authors.lastname: Paus
mesh_terms: D018093:Gelatinases / Q000378:metabolism*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007150:Immunohistochemistry; D008407:Mast Cells / Q000378:metabolism* / Q000473:pathology; D020778:Matrix Metalloproteinase 2; D008666:Metalloendopeptidases / Q000378:metabolism*; D012867:Skin / Q000378:metabolism* / Q000473:pathology; D012878:Skin Neoplasms / Q000378:metabolism / Q000473:pathology
chemical_list: D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2