Electron Microscopy of the Glomerular Basement Membrane of the Rat Kidney

2015 ◽  
pp. 45-53
Author(s):  
K. Oshima ◽  
M. Hatano ◽  
Y. Maeyama ◽  
N. Sugino ◽  
T. Takeuchi
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Rat Kidney

scholarly journals Anionic charge concentration of rat kidney glomeruli and glomerular basement membrane

1993 ◽  
Vol 289 (3) ◽  
pp. 647-652 ◽  
Author(s):  
W D Comper ◽  
A S N Lee ◽  
M Tay ◽  
Y Adal
Keyword(s):  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Electrostatic Interactions ◽  
Enzyme Inhibitors ◽  
Rat Kidney ◽  
Basement Membranes ◽  
Sulphate Content ◽  
Anionic Charge ◽  
Charge Concentration ◽  
A Charge

Estimates of levels of glomerular and glomerular-basement-membrane anion charge should serve as useful quantitative markers for the integrity of the tissues in health and disease. We have developed a simple, rapid, technique to measure this charge through the use of ion exchange with radioisotopes 22Na+ and 36Cl- at low ionic strengths in phosphate buffer. When this technique is used, normal glomeruli isolated from rat have a measured net anion charge concentration of 17.4 +/- 3.7 p-equiv. per glomerulus (n = 20). Perfused rat kidneys that lose approximately half of their glomerular heparan [35S]sulphate content (owing to oxygen-radical damage) exhibited a lower anion charge, of 7.5 +/- 1.6 p-equiv. per glomerulus (n = 5). Glomerular basement membranes prepared from rat glomeruli by a sonication-centrifugation procedure in the presence of enzyme inhibitors had a charge concentration of 6.3 +/- 0.7 mu-equiv./g wet wt. of tissue (n = 4), whereas membranes prepared by sonication, centrifugation, DNAse and detergent treatment had a charge concentration of 7.1 +/- 1.6 mu-equiv./g wet wt. (n = 4). Isotope-dilution experiments with 3H2O on these detergent-prepared glomerular basement membranes demonstrated that they had a water content of approx. 93%, which would then give a net anion charge concentration of 7.6 +/- 1.7 m-equiv./l (n = 4). These values are in good agreement with those obtained by others using titration techniques [Bray and Robinson (1984) Kidney Int. 25, 527-533]. The relatively low magnitude of glomerular anion charge in normal kidneys is consistent with other recent findings that glomerular anion charge is too low to affect the glomerular transport of charged molecules in a direct, passive, biophysical manner through electrostatic interactions.

scholarly journals Evaluation of ultrastructural alterations of glomerular basement membrane and podocytes in glomeruli by low-vacuum scanning electron microscopy

2021 ◽  
Author(s):  
Ping Lan ◽  
Dedong Kang ◽  
Akiko Mii ◽  
Yoko Endo ◽  
Masako Tagawa ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Assessment Tool ◽  
Periodic Acid ◽  
Renal Pathology ◽  
Blue Staining ◽  
Formalin Fixed Paraffin Embedded ◽  
Scanning Electron

Abstract Background Low-vacuum scanning electron microscopy (LV-SEM) is applied to diagnostic renal pathology. Methods To demonstrate the usefulness of LV-SEM and to clarify the optimal conditions of pathology samples, we investigated the alterations of glomerular basement membrane (GBM) and podocytes in control and experimental active Heymann nephritis (AHN) rats by LV-SEM. Results On week 15 following induction of AHN, spike formation on GBM with diffuse deposition of IgG and C3 developed. Using LV-SEM, diffuse crater-like protrusions were clearly noted three-dimensionally (3D) on surface of GBM in the same specimens of light microscopy (LM) and immunofluorescence (IF) studies only after removal coverslips or further adding periodic acid-silver methenamine (PAM) staining. These 3D ultrastructural findings of GBM surface could be detected in PAM-stained specimens by LV-SEM, although true GBM surface findings could not be obtained in acellular glomeruli, because some subepithelial deposits remained on surface of GBM. Adequate thickness was 1.5–5 μm for 10% formalin-fixed paraffin-embedded (FFPE) and 5–10 μm for the unfixed frozen sections. The foot processes and their effacement of podocytes could be observed by LV-SEM using 10%FFPE specimens with platinum blue (Pt-blue) staining or double staining of PAM and Pt-blue. These findings were obtained more large areas in 2.5% glutaraldehyde-fixed paraffin-embedded (2.5%GFPE) specimens. Conclusion Our findings suggest that LV-SEM is a useful assessment tool for evaluating the alterations of GBM and podocytes in renal pathology using routine LM and IF specimens, as well as 2.5%GFPE specimens.

Differentiation of glomerular filter and tubular reabsorption apparatus during foetal development of the rat kidney

Development ◽  
1980 ◽  
Vol 58 (1) ◽  
pp. 157-175
Author(s):  
Jean Schaeverbeke ◽  
Madeleine Cheignon
Keyword(s):  
Molecular Weight ◽  
Basement Membrane ◽  
Proximal Tubule ◽  
High Molecular Weight ◽  
Glomerular Basement Membrane ◽  
Rat Kidney ◽  
Tubular Reabsorption ◽  
Foetal Development ◽  
Endocytic Vesicles ◽  
Glomerular Barrier

Differentiation of the glomerulus and the proximal tubule was studied in the rat foetus, especially with regard to the development of the protein filtration-reabsorption apparatus. Filtration starts several days before full differentiation of the glomerulus, when the glomerular basement membrane consists of a thin lamina alongside the podocyte membrane. Endocytosis is functional from this time, but fusion between endocytic vesicles and lysosome-like bodies occurs 2 days later. Foetal urine electrophoresis shows the presence of many proteins, including high molecular weight ones, this proteinuria seeming chiefly due to the immaturity of the glomerular barrier.

Synthesis of the glomerular basement membrane in the rat kidney

1976 ◽  
Vol 20 (1) ◽  
pp. 125-137
Author(s):  
W. Romen ◽  
B. Schultze ◽  
K. Hempel
Keyword(s):  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Rat Kidney

scholarly journals Cells of renin lineage take on a podocyte phenotype in aging nephropathy

2014 ◽  
Vol 306 (10) ◽  
pp. F1198-F1209 ◽  
Author(s):  
Jeffrey W. Pippin ◽  
Sean T. Glenn ◽  
Ronald D. Krofft ◽  
Michael E. Rusiniak ◽  
Charles E. Alpers ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Basement Membrane ◽  
Progenitor Cells ◽  
Cell Fate ◽  
Glomerular Basement Membrane ◽  
Reporter Mice ◽  
Transmission Electron ◽  
Podocyte Proteins ◽  
Effects Of Aging

Aging nephropathy is characterized by podocyte depletion accompanied by progressive glomerulosclerosis. Replacement of terminally differentiated podocytes by local stem/progenitor cells is likely a critical mechanism for their regeneration. Recent studies have shown that cells of renin lineage (CoRL), normally restricted to the kidney's extraglomerular compartment, might serve this role after an abrupt depletion in podocyte number. To determine the effects of aging on the CoRL reserve and if CoRL moved from an extra- to the intraglomerular compartment during aging, genetic cell fate mapping was performed in aging Ren1cCre × Rs-ZsGreen reporter mice. Podocyte number decreased and glomerular scarring increased with advanced age. CoRL number decreased in the juxtaglomerular compartment with age. There was a paradoxical increase in CoRL in the intraglomerular compartment at 52 and 64 wk of age, where a subset coexpressed the podocyte proteins nephrin, podocin, and synaptopodin. Transmission electron microscopy studies showed that a subset of labeled CoRL in the glomerulus displayed foot processes, which attached to the glomerular basement membrane. No CoRL in the glomerular compartment stained for renin. These results suggest that, despite a decrease in the reserve, a subpopulation of CoRL moves to the glomerulus after chronic podocyte depletion in aging nephropathy, where they acquire a podocyte-like phenotype. This suggests that they might serve as adult podocyte stem/progenitor cells under these conditions, albeit in insufficient numbers to fully replace podocytes depleted with age.

scholarly journals The use of electron microscopy in the diagnosis of focal segmental glomerulosclerosis: are current pathological techniques missing important abnormalities in the glomerular basement membrane?

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1204
Author(s):  
Justin Davis ◽  
Alwie Tjipto ◽  
Katharine Hegerty ◽  
Andrew Mallett
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Focal Segmental Glomerulosclerosis ◽  
Glomerular Basement Membrane ◽  
Type Iv Collagen ◽  
Histological Diagnosis ◽  
Ultrastructural Changes ◽  
Type Iv ◽  
Segmental Glomerulosclerosis ◽  
Two Samples

Background: There is an increasing appreciation that variants of the COL4A genes may be associated with the development of focal segmental glomerulosclerosis (FSGS). On electron microscopy, such variants may produce characteristic changes within the glomerular basement membrane (GBM). These changes may be missed if glomerular lesions histologically diagnosed as FSGS on light microscopy are not subjected to electron microscopy. Methods: We conducted a retrospective cohort analysis of all patients presenting to two hospitals who received a primary histological diagnosis of FSGS to see if these samples underwent subsequent electron microscopy. Each such sample was also scrutinised for the presence of characteristic changes of an underlying type IV collagen disorder Results: A total of 43 patients were identified. Of these, only 30 underwent electron microscopy. In two samples there were histological changes detected that might have suggested the underlying presence of a type IV collagen disorder. Around one in three biopsy samples that had a histological diagnosis of FSGS were not subjected to electron microscopy. Conclusion: Renal biopsy samples that have a histological diagnosis of primary FSGS not subjected to subsequent electron microscopy may potentially miss ultrastructural changes in the GBM that could signify an underlying type IV collagen disorder as the patient’s underlying disease process. This could potentially affect both them and their families’ investigative and management decisions given potential for implications for transplant, heritability and different disease pathogenesis. This represents a gap in care which should be reflected upon and rectified via iterative standard care and unit-level quality assurance initiatives.

Alport Syndrome and Thin Glomerular Basement Membrane Nephropathy: A Practical Approach to Diagnosis

2009 ◽  
Vol 133 (2) ◽  
pp. 224-232 ◽  
Author(s):  
Mark Haas
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Renal Biopsy ◽  
Glomerular Basement Membrane ◽  
Alport Syndrome ◽  
Autosomal Recessive ◽  
Type Iv Collagen ◽  
The Past ◽  
Type Iv ◽  
Pathologic Features

Abstract Context.—Alport syndrome and thin glomerular basement membrane nephropathy (TBMN) are genetically heterogenous conditions characterized by structural abnormalities in the glomerular basement membrane and an initial presentation that usually involves hematuria. Approximately 40% of patients with TBMN are heterozygous carriers for autosomal recessive Alport syndrome, with mutations at the genetic locus encoding type IV collagen α3 [α3(IV)] and α4 chains. However, although the clinical course of TBMN is usually benign, Alport syndrome, particularly the X-linked form with mutations in the locus encoding the α5 chain of type IV collagen [α5(IV)], typically results in end-stage renal disease. Electron microscopy is essential to diagnosis of TBMN and Alport syndrome on renal biopsy, although electron microscopy alone is of limited value in distinguishing between TBMN, the heterozygous carrier state of X-linked Alport syndrome, autosomal recessive Alport syndrome, and even early stages of X-linked Alport syndrome. Objectives.—To review diagnostic pathologic features of each of the above conditions, emphasizing the need for immunohistology for α3(IV) and α5(IV) in addition to electron microscopy to resolve this differential diagnosis on a renal biopsy. The diagnostic value of immunofluorescence studies for α5(IV) on a skin biopsy in family members of patients with Alport syndrome also is reviewed. Data Sources.—Original and comprehensive review articles on the diagnosis of Alport syndrome and TBMN from the past 35 years, primarily the past 2 decades, and experience in our own renal pathology laboratory. Conclusions.—Although Alport syndrome variants and TBMN do not show characteristic light microscopic findings and can be difficult to differentiate from each other even by electron microscopy, using a combination of electron microscopy and immunohistology for α3(IV) and α5(IV) enables pathologists to definitively diagnose these disorders on renal biopsy in most cases.

Thin Glomerular Basement Membrane Nephropathy: Incidence in 3471 Consecutive Renal Biopsies Examined by Electron Microscopy

2006 ◽  
Vol 130 (5) ◽  
pp. 699-706 ◽  
Author(s):  
Mark Haas
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Glomerular Basement Membrane ◽  
Medical Center ◽  
Academic Medical Center ◽  
Minimal Change ◽  
Minimal Change Nephropathy ◽  
Normal Ranges ◽  
Renal Biopsies ◽  
20 Nm

Abstract Context.—Thin glomerular basement membrane (GBM) nephropathy is often equated with benign familial hematuria, although it may occur sporadically and may not always be benign. Thin GBM nephropathy is reported to occur in at least 1% of the population, although its reported incidence varies considerably in different studies because there are presently no uniform criteria for its diagnosis by electron microscopy (EM). Objective.—To determine the incidence of thin GBM nephropathy in a large sample of renal biopsies using a basic methodology that can easily be applied in any diagnostic EM laboratory. Design.—Direct measurements of GBM thickness were made from electron micrographs at 3 specified points along each of 10 randomly selected glomerular capillaries to determine an average GBM thickness for each of 50 males and 50 females, ages 9 to 80 years, with minimal-change nephropathy or acute interstitial nephritis, without hematuria. The means of the average GBM thickness values were 330 ± 50 (SD) nm in the males and 305 ± 45 nm in the females; normal ranges for each sex were defined as being within 2 SD of these means. The average GBM thickness was then similarly determined for renal biopsies with suspected thin GBMs examined from January 2000 to December 2004; a total of 3471 renal biopsies were examined by EM during this period. Setting.—Academic medical center renal pathology/EM laboratory. Results.—Excluding biopsies with immunoglobulin A nephropathy, which is known to be frequently associated with thin GBMs, and biopsies with Alport syndrome, 67 biopsies (1.9% of total) had an average GBM thickness below the sex-specific normal range. Of these, 37 biopsies were performed specifically because of hematuria and had an average GBM thickness of 121 to 215 nm (mean, 185 ± 20 nm). The remaining 30 (0.9%) biopsies, with average GBM thicknesses of 143 to 227 nm (mean, 190 ± 20 nm), represent cases of incidentally discovered thin GBM nephropathy. Conclusions.—Based on the frequency of incidentally discovered cases and taking into account excluded cases and biopsies (eg, with diabetic nephropathy) in which diagnosis of incidental thin GBM nephropathy by EM is not possible, the incidence of thin GBM nephropathy in our population is estimated to be between 1% and 2%. Diseases most often associated with incidental thin GBM nephropathy were focal segmental glomerulosclerosis (10 cases) and minimal-change nephropathy (5 cases).

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