[Detections of matrix metalloproteinases activities and localization by film in situ zymography (FIZ)].
Extracellular proteolysis is an essential process for cell migration in several diseases such as tumor invasion, metastasis, and angiogenesis. In an invasive process, matrix metalloproteinases (MMPs) play a central role as degradation enzymes of extracellular matrix. Immunohistochemical staining (IHC) or ELISA methods have been carried out for the detection of tissue MMP proteins, and MMP activities have been mainly measured by gelatin zymography. Recently, film in situ zymography (FIZ) was developed for the regional detection of tissue MMP activities. FIZ uses a polyester film coated with gelatin uniformly and thinly. Frozen sections are incubated on the film at 37 degrees for optimal time. After gelatin staining with Biebrich Scarlet, the unstained area corresponds to MMP activities. The regional detection of tissue MMP activities by FIZ is simple and quantitative, and it is a useful tool for the studies of many diseases involving MMP. The detection of precise localization of MMP activity in tissues by FIZ may contribute the new classification of diseases involving MMP.
Muramatsu
Michiko
M
Department of Pharmacology, Osaka Medical College, Takatsuki City, Osaka 569-8686, Japan.
Takai
Shinji
S
Miyazaki
Mizuo
M
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D053818:Chymases; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005780:Gelatin; D020782:Matrix Metalloproteinases / Q000032:analysis* / Q000502:physiology; D023421:Models, Animal; D018919:Neovascularization, Physiologic / Q000502:physiology; D012697:Serine Endopeptidases / Q000502:physiology
D015415:Biomarkers; C015233:gelatin film; D005780:Gelatin; D012697:Serine Endopeptidases; D053818:Chymases; D020782:Matrix Metalloproteinases