Entactin/nidogen (E/N) was isolated from bovine renal tubular basement membrane. Apparent molecular weight, amino acid composition, and molecular configuration by electron microscopy rotary shadowing were similar to that of nidogen from EHS mouse tumor. The identity of bovine E/N was confirmed using a thrombin derived peptide, the sequence of which corresponded to a region within mouse and human E/N. Monoclonal and polyclonal anti-E/N antibodies were used to determine the distribution of E/N in human kidney by immunofluorescent and immunoelectron microscopy. E/N was present in all renal basement membranes and was distributed through the full width of the glomerular basement membrane (GBM) with accentuation along its epithelial aspects. E/N distribution was similar to that of novel collagen chain α3(IV) NC domain in the GBM. In the mesangium, E/N was distributed mainly in the peripheral mesangial region that is bounded by the GBM, while classical collagen chain α1(IV) NC as present diffusely throughout the mesangium. In the developing nephron, E/N was present in basement membranes of the ureteric bud, primitive vesicle and S-form. In all instances, E/N co-localized with laminin B2 chain. Prominent E/N detection within the mesangium was observed in diseases where mesangial expansion was present. This process was also seen in early diabetic nephropathy, but disappeared with disease progression. However, all thickened diabetic renal basement membranes showed an increase in E/N which was also present in Kimmelstiel-Wilson lesions. E/N was observed in the GBM "spikes" of membranous glomerulonephritis and in epithelial crescents associated with various disorders. The association between E/N, laminin and type IV collagen chains observed in the normal kidney were maintained in disorders with altered E/N distribution. We could not detect any changes in the distribution of E/N in other acquired and hereditary kidney diseases. These observations reflect the involvement of E/N in the structure and disease alteration of renal basement membranes and mesangial matrix.
Bibliographical noteFunding Information:
DKO 07087, and DK 36007, The National Kidney Foundation of the Upper Midwest, The Juvenile Diabetes Foundation International, and the Viking Children's Fund. EHS nidogen used in this study was provided by Dr. Aris Charonis, University of Minnesota, and rabbit antiserum to mouse tumor EHS-derived nidogen was from R. Timpl, Max Planck Institute for Biochemistry. The authors gratefully acknowl- edge the help and assistance of the following individuals in this work: Vickie VanDrisse, (laboratory studies), Thomas Groppoli (immunoelec- (ron microscopy), Crystal Blocher and Kathy Divine (immunohistol-ogy), Drs. E. Tsilibary and A. Charonis (rotary shadowing), Marshall Hoff (illustrations), and Patricia Erickson (manuscript preparation).