title: Expression of gelatinase A, a mediator of extracellular matrix remodeling, by tracheal gland serous cells in culture and in vivo.
abstract: Tracheal gland morphogenesis and gland hypertrophy in disease involve the penetration of epithelial cells into the submucosa, a process that requires digestion of the basal lamina and the surrounding extracellular matrix. We observed that bovine tracheal gland cells invaded collagen substrates and were inhibited from doing so in the presence of a metalloproteinase inhibitor GM6001. The gland cells, but not bovine tracheal surface epithelial cells, secreted a 72-kDa metalloproteinase. The purified enzyme could be activated with 4-aminophenylmercuric acetate and converted to an active 65-kDa form that was far more effective in degrading denatured collagen (gelatin) than nondenatured type I and IV collagens and was ineffective in degrading intact interstitial collagen fibers. At 25 degrees C, the initial rate of degradation of acid-solubilized type I collagen was approximately 50 mg of type I collagen cleaved per min per mg of enzyme, whereas acid-solubilized type IV collagen was degraded at approximately 250 mg cleaved per min per mg of enzyme. In contrast, at the same temperature, heat-denatured type I collagen was degraded 1000-fold more rapidly, while heat-denatured type IV collagen was cleaved 50-fold more rapidly. The activity of the enzyme was maximal at pH 7-8 and was completely abolished by the metalloproteinase inhibitors EDTA and 1,10-phenanthroline. In immunoblots, the enzyme was recognized by an antibody directed against human gelatinase A, the 72-kDa gelatinase. The purified enzyme disrupted the distribution pattern of type IV collagen in the gland basal lamina, as well as of interstitial collagen in the underlying stromal tissue, as shown in tissue sections by immunocytochemistry. Using an antibody directed against the purified enzyme, we also showed by immunocytochemistry that the gelatinase was present in tracheal tissue and was specifically located at the periphery of some tracheal gland acini. Northern blots showed higher concentrations of gelatinase A mRNA in glands than in epithelium microdissected from adult cow tracheas. These data indicate that gelatinase A is a specialized product of the tracheal gland epithelial cell, a cell type normally invasive as part of its developmental program; the enzyme may play an important role in normal gland development and disease-associated hypertrophy.
authors.affiliation: Department of Anatomy, University of California, San Francisco 94143-0452.
authors.forename: J M
authors.initials: JM
authors.lastname: Tournier
authors.forename: M
authors.initials: M
authors.lastname: Polette
authors.forename: J
authors.initials: J
authors.lastname: Hinnrasky
authors.forename: J
authors.initials: J
authors.lastname: Beck
authors.forename: Z
authors.initials: Z
authors.lastname: Werb
authors.forename: C
authors.initials: C
authors.lastname: Basbaum
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000378:metabolism; D015152:Blotting, Northern; D015153:Blotting, Western; D002417:Cattle; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003238:Connective Tissue / Q000378:metabolism; D003429:Cross Reactions; D004848:Epithelium; D005088:Exocrine Glands / Q000166:cytology / Q000201:enzymology*; D005109:Extracellular Matrix / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D018093:Gelatinases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D016591:Histocytological Preparation Techniques; D020778:Matrix Metalloproteinase 2; D008666:Metalloendopeptidases / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D012704:Serous Membrane / Q000166:cytology / Q000201:enzymology*; D014132:Trachea / Q000166:cytology / Q000201:enzymology*
chemical_list: D003094:Collagen; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2