title: Use of gel zymography to examine matrix metalloproteinase (gelatinase) expression in brain tissue or in primary glial cultures.
abstract: Glia synthesize, package, and secrete several species of matrix proteases, including the gelatinases (pro-)MMP-2 and (pro-)MMP-9. In appropriate settings (e.g., experimental ischemia), these MMPs can be assayed from cerebral tissues or from astrocytes and microglia in culture by enzymatic substrate-dependent assays and by gelatin-based zymography. We describe the methodologies for the sensitive quantitative development of the inactive and active forms of both MMP-2 and MMP-9 from tissues and cells, by means of lysis of the collagen substrate in collagen-impregnated gel electropheresis by the zymogen and active gelatinases. These methodologies are a refinement of those used commonly, with instructions to increase sensitivity. Serious and often overlooked issues regarding sources of sample contamination and elements confounding the MMP band development and their interpretation are discussed.
authors.affiliation: Department of Pediatrics, University of California San Diego, San Diego, CA, USA.
authors.forename: Harald
authors.initials: H
authors.lastname: Frankowski
authors.forename: Yu-Huan
authors.initials: YH
authors.lastname: Gu
authors.forename: Ji Hoe
authors.initials: JH
authors.lastname: Heo
authors.forename: Richard
authors.initials: R
authors.lastname: Milner
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Del Zoppo
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002478:Cells, Cultured; D003094:Collagen; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007511:Ischemia / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D051379:Mice; D009457:Neuroglia / Q000378:metabolism*
chemical_list: D003094:Collagen; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9