Matrix metalloproteinases and TIMPS in cultured C57BL/6J-cpk kidney tubules.
Restructuring of basement membranes is a hallmark of the pathology of renal cystic disorders. Here, we present findings consistent with the view that basement membrane degradation by matrix metallo-proteinases (MMPs) may contribute to abnormal basement membrane structure in polycystic kidney disease. Cells from cystic kidney tubules embedded in collagen gels appeared to migrate through the gel. This migration through collagen indicated that these cells could degrade the matrix. To examine this activity, we cultured cystic kidney tubules derived from the C57BL/6J cpk/cpk mouse, a hereditary model of polycystic kidney disease, and assayed conditioned medium for the presence of MMPs and tissue inhibitors of metalloproteinases (TIMPs). The conditioned medium from the cystic tubules contained higher than normal levels of MMP-9, MMP-2, and MMP-3 as well as TIMP-1 and TIMP-2. A 101 kDa protease was present equally in cystic and control cultures and although inhibited by EDTA, it was not inhibited by TIMPs, nor activated by the mercurial compound APMA. These data suggest that cystic kidney tubules synthesize and secrete high levels of MMPs which may then participate in the restructuring of the tubular basement membrane.
Rankin
C A
CA
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, USA.
Suzuki
K
K
Itoh
Y
Y
Ziemer
D M
DM
Grantham
J J
JJ
Calvet
J P
JP
Nagase
H
H
D000818:Animals; D001485:Basement Membrane / Q000201:enzymology / Q000473:pathology; D015152:Blotting, Northern; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D017364:Collagenases / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D018093:Gelatinases / Q000235:genetics / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D052177:Kidney Diseases, Cystic / Q000201:enzymology*; D007684:Kidney Tubules / Q000166:cytology*; D008297:Male; D020778:Matrix Metalloproteinase 2; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives / Q000494:pharmacology; D011480:Protease Inhibitors / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D019716:Tissue Inhibitor of Metalloproteinase-2; D019714:Tissue Inhibitor of Metalloproteinases
D017077:Culture Media, Conditioned; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D011480:Protease Inhibitors; D011506:Proteins; D012333:RNA, Messenger; D013439:Sulfhydryl Reagents; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; C010024:4-aminophenylmercuriacetate; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D010662:Phenylmercuric Acetate