Further observations on the morphological alterations of the glomerular capillary wall of the rat kidney caused by chemical and physical agents: standard procedures versus quick-freezing and freeze-substitution

The purpose of the present study was to pin-point which of the various preparatory steps required by different histological procedures (conventional, cryosubstitution, freezing followed by cryosubstitution or replication) induces alterations in the fine structure of the endothelial and epithelial su...

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Bibliographic Details
Published in:The Histochemical journal 1993-05, Vol.25 (5), p.357-366
Main Authors: Reale, E, Luciano, L
Format: Article
Language:English
Subjects:
Acetone
Animals
Basement Membrane - drug effects
Basement Membrane - ultrastructure
Capillaries - ultrastructure
Cryopreservation
Desiccation
Endothelium, Vascular - ultrastructure
Epithelium - ultrastructure
Ethanol
Freeze Substitution
Glutaral
Kidney Glomerulus - blood supply
Kidney Glomerulus - ultrastructure
Male
Osmium Tetroxide
Rats
Rats, Wistar
Tissue Embedding
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Summary:The purpose of the present study was to pin-point which of the various preparatory steps required by different histological procedures (conventional, cryosubstitution, freezing followed by cryosubstitution or replication) induces alterations in the fine structure of the endothelial and epithelial surface coat (SC) and of the interposed glomerular basement membrane (GBM). Samples of rat kidney cortex prefixed by vascular perfusion, and isolated glomeruli fixed by immersion, were used. The results demonstrate a continuous, homogeneous and amorphous SC and a GBM devoid of laminae rarae when freezing is used and followed either by cryosubstitution and embedding or by fracture, deep etching and replication. Postfixation in OsO4, and especially dehydration in organic solvents at room temperature, generate a filamentous SC and a GBM with laminae rarae. The different morphology may be due to an extraction of glycoconjugates from SC and GBM during postosmication and/or especially to precipitation of these components during dehydration by organic solvents at room temperature. Studies on isolated glomeruli show comparable results and, in addition, demonstrate that the distance of the capillary loop from the surface of the block influences the morphology of SC and GBM. Rinsing the vascular bed before isolation does not induce a loss of SC or GBM components.
ISSN:0018-2214
1573-6865
DOI:10.1007/BF00159500