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An ultrastructural analysis of isolated basement membranes in the acellular renal cortex: A comparative study of human and laboratory animals
Authors:Edward C Carlson  M Cristina Kenney
Abstract:Freshly harvested kidneys from New Zealand white rabbits, Sprague-Dawley white rats, rhesus monkeys, and transplant-quality human kidneys were used in this study. Minced renal cortical tissue blocks (<2 mm3) were treated with 1 mM EDTA, 3% Triton X-100, 0.025% DNAse, and 4% sodium deoxycholate in an effort to remove all cellular elements and leave the extracellular matrix (ECM) intact. These preparations showed remarkable structural preservation and all components of the ECM, including basement membranes (BMs), maintained their in vivo histoarchitectural relationships. By light microscopy, at least four major BM types were recognizable, including Bowman's capsular BM (BCBM), tubular BM (TBM), glomerular BM (GBM), and peritubular capillary BM (PTCBM). Scanning electron microscopy demonstrated that, despite the lack of supporting interstitium, GBMs in human, monkey, and rat (and rabbit to a lesser degree) exhibit intrinsic structural rigidity such that their convoluted spheroidal shapes are maintained following cell removal. Transmission electron microscopy showed that major BM types are morphologically heterogeneous and vary markedly within and between species. Randomized measurements showed that isolated BM thicknesses (lamina densa only) compared favorably with those reported in cellular preparations. Mean thicknesses of GBMs were within normal ranges in all species with or without power transformations to reduce right-sided skew of distribution curves. In all species, thickness of BCBM > TBM > GGBM > PTCBM. The striking morphologic heterogeneity of major BM types demonstrated in the acellular renal cortex is not surprising in view of recent biochemical analyses that show that BMs derived from different sources are compositionally disparate. We conclude that BMs should be evaluated and characterized individually and that morphologic definition of isolated BMs is necessary prior to further analysis.
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