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A novel in‐situ ‐zymography technique localizes gelatinolytic activity in human skin to mast cells
Author(s) -
KrejciPapa N. C.,
Paus R.
Publication year - 1998
Publication title -
experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.108
H-Index - 96
eISSN - 1600-0625
pISSN - 0906-6705
DOI - 10.1111/j.1600-0625.1998.tb00331.x
Subject(s) - extracellular matrix , immunohistochemistry , matrix metalloproteinase , staining , mast cell , zymography , chemistry , proteases , microbiology and biotechnology , tryptase , biology , pathology , biochemistry , enzyme , immunology , medicine
Matrix‐metallo‐proteinases play a key role in cutaneous tissue remodeling and wound healing, and have been implicated as the ratelimiting 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 μm 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.