title: Role of the matrixin MMP-2 in multicellular organization of adipocytes cultured in basement membrane components.
abstract: Primary rat adipocytes cultured in basement membrane component gels migrated and organized into large, three-dimensional, multicellular clusters. Gross morphological changes seen during this reorganization are described. The rate of cluster formation decreased with age of the rats and was stimulated by insulin in older, but not in younger rats. Echistatin, a disintegrin, partially inhibited the formation of multicellular clusters in a concentration-dependent fashion (50% inhibitory concentration approximately 10 nM). The original extracellular matrix was initially remodeled and eventually destroyed by the time large multicellular clusters were observed. This implied that one or more matrix-degrading protease(s) were being secreted. Adipocyte-conditioned medium was found to contain a divalent cation-sensitive gelatinase activity at approximately 72 and/or approximately 62 kDa. The elution profile of this activity from gelatin-Sepharose 4B was similar to matrix metalloproteinase 2 (MMP-2, a 72-kDa matrixin with a 62-kDa mature form), and the dimethyl sulfoxide eluant from these columns contained MMP-2 immunoreactivity. MMP-2 concentration and activity were greater in conditioned medium from young than from older animals; however, insulin did not affect the amount of MMP-2 in adipocyte-conditioned media. The matrixin inhibitor 1,10-phenanthroline not only blocked gelatinase activity in zymograms but also prevented extracellular matrix remodeling and destruction, as well as adipocyte migration and the formation of cell-cell contacts in adipocyte cultures. These observations are consistent with the hypothesis that the matrixin MMP-2 is secreted by adipocytes. Whereas matrixin activity alone may not be sufficient for the formation of multicellular clusters, the data indicate that it may have a requisite role in this process.
authors.affiliation: Department of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey 17033, USA.
authors.forename: L M
authors.initials: LM
authors.lastname: Brown
authors.forename: H L
authors.initials: HL
authors.lastname: Fox
authors.forename: S A
authors.initials: SA
authors.lastname: Hazen
authors.forename: K F
authors.initials: KF
authors.lastname: LaNoue
authors.forename: S R
authors.initials: SR
authors.lastname: Rannels
authors.forename: C J
authors.initials: CJ
authors.lastname: Lynch
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D000367:Age Factors; D000818:Animals; D001485:Basement Membrane / Q000166:cytology; D002256:Carbonic Anhydrases / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002448:Cell Adhesion; D002449:Cell Aggregation / Q000187:drug effects; D002465:Cell Movement; D003094:Collagen; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins / Q000502:physiology; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D018093:Gelatinases / Q000502:physiology*; D007328:Insulin / Q000494:pharmacology; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D008297:Male; D020778:Matrix Metalloproteinase 2; D008666:Metalloendopeptidases / Q000502:physiology*; D018996:Myelin P2 Protein / Q000378:metabolism; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D010455:Peptides / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D011509:Proteoglycans; D051381:Rats; D017207:Rats, Sprague-Dawley; D019030:Receptors, Vitronectin / Q000037:antagonists & inhibitors
chemical_list: D002352:Carrier Proteins; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins; C490174:Fabp7 protein, rat; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D018996:Myelin P2 Protein; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D010455:Peptides; D010618:Phenanthrolines; D011509:Proteoglycans; D019030:Receptors, Vitronectin; C057986:echistatin; C059485:matrigel; D003094:Collagen; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D002256:Carbonic Anhydrases; C025205:1,10-phenanthroline