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Glomerular Basement Membrane

Q: What are the main components of the GBM?

The GBM is primarily composed of collagen IV, laminin, and other extracellular matrix proteins. These components work together to provide structural integrity and selective permeability, allowing the passage of small molecules while retaining larger proteins. Alterations to the GBM's composition and ultrastructure are implicated in various kidney diseases.

The glomerular basement membrade (GBM) is a specialized extracellular matrix structure that serves as the primary filtration barrier in the kidnye's glomeruli.
It is composed of collagen IV, laminin, and other components that provide structural support and selectively permit the passage of small molecules while retaining larger proteins.
Alterations to the GBM's composition and ultrastructure are implicated in various kidney diseases, making it an important target for research into the underlying mechanisms and potential therapies.
PubCompare.ai's AI-driven tools can help streamline your studies of the GBM by locating relevant protocols from the literature, preprints, and patents, and identifying the most reproducible and accurate research methodolgies.

Most cited protocols related to «Glomerular Basement Membrane»

Quantitative Kidney Biopsy Analysis

A subset of subjects underwent percutaneous kidney biopsy at the end of the treatment period to determine whether treatment with losartan was associated with structural differences. Baseline kidney biopsies were not performed because of safety concerns. Tissue was processed and embedded in epoxy resin (Epon 812) and prepared for microscopy as described previously (12 (link)). Light and electron microscopy were performed either in the Beckman Center for Electron Microscopy at Stanford University or in the Division of Nephrology at the University of Minnesota. Digital light and electron micrographs were used to make measurements using formal stereologic methods to account for two-dimensional sampling of three-dimensional objects (13 ). Predefined morphometric variables included glomerular volume, percent globally sclerotic glomeruli, fractional interstitial area, mesangial fractional volume, filtration surface area density, glomerular basement membrane width, number of endothelial cells, mesangial cells and podocytes per glomerulus, filtration slit frequency, and foot process width (12 (link),14 (link)–16 (link)).

Weil E.J., Fufaa G., Jones L.I., Lovato T., Lemley K.V., Hanson R.L., Knowler W.C., Bennett P.H., Yee B., Myers B.D, & Nelson R.G. (2013). Effect of Losartan on Prevention and Progression of Early Diabetic Nephropathy in American Indians With Type 2 Diabetes. Diabetes, 62(9), 3224-3231.

Publication 2013

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Evaluating siRNA Nanoparticle Stability

NPs were loaded with Forster Resonance Energy Transfer (FRET, using FAM and Cy5) pair-labeled 23mer dsDNAs (a model for siRNA) (FRET-NPs). Fluorescent intensity was measured using a spectrophotofluorometer with an excitation wavelength of 488nm (Jobin Yvon/Horiba Fluorolog-3 FL3-111, Horiba Scientific, Kyoto Japan). FAM emission was collected at 520nm ± 3nm, and Cy5 emission was obtained at 670nm ± 3nm. %FRET was calculated as a ratio of the fluorescent intensity as follows:

% F R E T = \frac{I_{670}}{I_{520} + I_{670}} .

For serum stability measurements, FRET-NPs were added into human whole blood diluted 1:3 in PBS at 100 nM (50 nM for each DNA). Treated blood samples were loaded into a black, round bottom 96 well plate and placed on a shaker for 5 minutes before incubating at 37°C for 1 h. Plates were then centrifuged at 500 × g for 5 minutes, and then 50 μL of supernatant (diluted blood serum) from each well was transferred into a black, clear bottom 96 well plate. Fluorescence was measured using a Microplate Reader and %FRET was calculated using Eqn 1. In parallel experiments to assess hemocompatibility ex vivo, polyplex NPs loaded with FAM-labeled dsDNA were used to quantify the percent of NPs in the supernatant, as a measure of inertness, or ability to reduce nonspecific adsorption to or aggregation with RBCs.
Because siRNA decomplexation by heparan sulfate-containing glomerular basement membrane (GBM) in the kidney is a primary cause for rapid systemic clearance of polycation-siRNA nanoparticles,^{39 , 40 (link)} the stability of FRET-NPs was measured in the presence of 2 U/mL of heparin sodium salt in DPBS. The fluorescence emission was measured over time using a microplate reader with an excitation wavelength of 488 nm and an emission wavelength of 670 nm (Tecan Infinite F500, Mannedorf, Switzerland).

Nelson C.E., Kintzing J.R., Hanna A., Shannon J.M., Gupta M.K, & Duvall C.L. (2013). Balancing Cationic and Hydrophobic Content of PEGylated siRNA Polyplexes Enhances Endosome Escape, Stability, Blood Circulation Time, and Bioactivity In Vivo. ACS nano, 7(10), 10.1021/nn403325f.

Publication 2013

Adsorption BLOOD DNA, Double-Stranded Erythrocytes Fluorescence Fluorescence Resonance Energy Transfer Glomerular Basement Membrane Heparin Sodium Homo sapiens Kidney polycations RNA, Small Interfering SALL2 protein, human Serum Sodium Chloride, Dietary Sulfate, Heparan

Histological and Immunohistochemical Analysis of Renal Fibrosis

Kidneys were fixed with phosphate buffered saline containing 4% paraformaldehyde for overnight and then embedded in paraffin. Sections (4 µm) were cut and deparaffinized in xylene, followed by rehydration in a graded series of ethanol. Staining was performed using hematoxylin and eosin, periodic acid Schiff (PAS), and Sirius red staining. Immunohistochemical staining was performed as described previously [24 (link)], using type I collagen (Abcam, Cambridge, UK), fibronectin (BD Biosciences, San Jose, CA, USA), β-tubulin (Abcam), phospho-Smad3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), TGF-β (Santa Cruz Biotechnology), and insulin (Santa Cruz Biotechnology). Thickness of the glomerular basement membrane (GBM) was measured by electron microscopy (H-7100; Hitachi, Tokyo, Japan). Renal fibrotic areas were quantified by morphometric analysis using a light microscope equipped with an imaging system containing aMRc5 Carl Zeiss microscope (Oberkochen, Germany) and iSolution DT version 7.7 software (IMT i-Solution, Coquitlam, BC, Canada). Areas of positive PAS matrix, Sirius red, immunostaining for type I collagen, and fibronectin in the renal fibrotic regions (brown color) were quantified by computer-based morphometric analysis. All data were normalized to the control and expressed as fold increase relative to the control.

Jung G.S., Jeon J.H., Choe M.S., Kim S.W., Lee I.K., Kim M.K, & Park K.G. (2016). Renoprotective Effect of Gemigliptin, a Dipeptidyl Peptidase-4 Inhibitor, in Streptozotocin-Induced Type 1 Diabetic Mice. Diabetes & Metabolism Journal, 40(3), 211-221.

Publication 2016

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Detailed Kidney Histopathology Assessment

Kidney specimens were processed by light and immunofluorescence microscopic examination. For the light microscopy, the right kidneys from each animal were fixed in 10% phosphate-buffered formalin solution and embedded in paraffin. Sections of 2 μm thickness were cut and stained with hematoxylin and eosin (HE) and periodic acid-Schiff (PAS). To evaluate the glomerular hypercellularity, at least 10 glomeruli were examined for each animal, the number of cells in each glomeruli (including endothelial cells, mesangial cells, and podocytes) was counted and average number was calculated; meanwhile, the incidence of glomerular basement membrane thickening or mesangial proliferation among 100 glomeruli was calculated too.
To assess the tubulointerstitial damage, a semiquantitative method of renal histology using a grading scale of 0–4 was applied: 0: normal; 1: lesions in <25% of the area; 2: lesions in 25% to 50% of the area; 3: lesions in >50% of the area; and 4: lesions involving the entire area [9 (link), 10 (link)]. Tubular atrophy, dilation, casts, interstitial inflammation, and fibrosis were assessed in 10 kidney fields at a magnification of ×100.
For immunofluorescence microscopy, tissue blocks from the left kidney were instantaneously frozen in n-hexane precooled to −70°C, and 4 μm cryostat sections were stained with fluorescein isothiocyanate- (FITC-) conjugated anti-rat IgG. The degree of deposition of immune complex was calculated quantitatively on the basis of the staining intensity and distribution.

Zhang S., Xin H., Li Y., Zhang D., Shi J., Yang J, & Chen X. (2013). Skimmin, a Coumarin from Hydrangea paniculata, Slows down the Progression of Membranous Glomerulonephritis by Anti-Inflammatory Effects and Inhibiting Immune Complex Deposition. Evidence-based Complementary and Alternative Medicine : eCAM, 2013, 819296.

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Publication 2013

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Quantitative Ultrastructural Analysis of Aged Kidneys

A wedge biopsy (>15 mm in length) was taken from the outer cortex of each donated kidney immediately before transplant and fixed in a paraformaldehyde/gluteraldehyde solution and embedded in LX-112 epoxy resin (Ladd Research, Williston, VT, USA). Tissue was prepared for light microscopy and electron microscopy as described previously.^{2 (link)}The prevalence of global sclerosis was determined in the × 50 light microscopic images using an equation that takes into account the smaller diameter of sclerotic glomeruli, and the consequent difference in the probability of encountering a glomerulus of either type in a random cross-section.^{40 (link),41 (link)} Fractional interstitial area and glomerular volume were determined by light microscopy images.^{2 (link)} The mean number (±1 s.d.) of glomeruli available for determination of the tuft areas was 47±41 and 60±40 in younger and older groups, respectively. Montages of three whole glomerular profiles were prepared from the × 3000 transmission electron microscopy images for determination of filtration surface density (S_v) of the peripheral glomerular capillary wall by line-intercept methods. Filtration surface area provided by the peripheral capillary wall per glomerulus (S) was calculated from the product of S_v and glomerular volume;⁴² we used resin-embedded sections to circumvent the substantial glomerular shrinkage associated with paraffin embedding.^{43 (link)}The assessment of hydraulic permeability (k) of the glomerular capillary walls was performed at the ultrastructural level ( × 12,000). Dimensions of the podocyte layer that accounts for 50% resistance to water flow included the width and the frequency of the filtration slits, in which the latter is determined by counting the total number of epithelial filtration slits and dividing it by the total length of the peripheral glomerular capillary wall.^{40 (link),41 (link)} The glomerular basement membrane accounts for the remaining resistance to water flow, and its thickness, an important determinant of k, was calculated from the harmonic mean glomerular basement membrane thickness as measured by the orthogonal intercept method.^{44 (link)} SNK_f was then calculated from the product of S and k using an ultrastructural-hydrodynamic model of viscous flow as described by us previously.^{40 (link),41 (link)} Finally, we computed the N_FG per kidney, where
Because of assumptions used in the calculation of both whole-kidney and SNK_f values, the calculated value of N_FG should be regarded as an approximation only.

Tan J.C., Busque S., Workeneh B., Ho B., Derby G., Blouch K.L., Sommer F.G., Edwards B, & Myers B.D. (2010). Effects of aging on glomerular function and number in living kidney donors. Kidney International, 78(7), 686-692.

Publication 2010

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Most recents protocols related to «Glomerular Basement Membrane»

Quantitative Analysis of Kidney Histology

Kidneys were harvested at 2, 6, or 12 weeks. Formalin-fixed, paraffin-embedded kidneys were sectioned at 4 μm thickness. PAS-stained sections were scanned at 200× magnification using Aperio ImageScope from Leica Biosystems, and image analysis was performed using QuPath software (v0.2.2). Glomerular surface area was measured manually by a blinded investigator by outlining the entire glomerular basement membrane area of the glomerular globe, excluding Bowman’s capsule, and using QuPath software to measure surface area in the measured plane. All glomeruli in the scanned kidney section were measured (range 50–200 glomeruli per kidney) to reduce sampling bias. The narrowest proximal tubular profiles in the outer strip of the outer medulla were selected, and diameter was manually measured. In total, 50 tubules in half kidney sections and 100 tubules in entire kidney sections were measured. All measurements were performed by an investigator blinded to mouse genotypes and treatment. Tubular size and glomerular surface area were analyzed using mixed-effects regression models accounting for within-subject correlations of tubular diameter or log glomerular surface area. All mean values reported for glomerular surface are geometric means.

Good P.I., Li L., Hurst H.A., Serrano Herrera I., Xu K., Rao M., Bateman D.A., Al-Awqati Q., D’Agati V.D., Costantini F, & Lin F. (2023). Low nephron endowment increases susceptibility to renal stress and chronic kidney disease. JCI Insight, 8(3), e161316.

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Publication 2023

Bowmans Capsule Eye Formalin Genotype Glomerular Basement Membrane Kidney Kidney Glomerulus Medulla Oblongata Mus Paraffin Tubule, Kidney

Ultrastructural Analysis of Glomerular Endothelium

The kidney specimens were fixed with a 4% paraformaldehyde and then incubated with 1% osmium tetroxide (OsO₄) at 4 °C for 1 h. The fixed specimens were dehydrated with gradient alcohols and embedded in Epon. Next, 70–100 nm ultrathin sections were cut with a Leica EM UC 7 microtome and stained with 1% uranyl acetate and lead citrate. Electron micrographs were taken at 30.000× at 80 kV with a JEM 1400 electron microscope (JEOL, Tokyo, Japan) and analyzed using ImageJ software (National Institutes of Health, Bethesda, MD, USA). Microscopic analysis was carried out at the Multiple-access Center for Microscopy of Biological Subjects (Institute of Cytology and Genetics, Novosibirsk, Russia). The number of fenestrae of endotheliocytes of glomerular capillaries and the number of podocyte foot processes were determined for 2 μm of the glomerular basement membrane. Additionally, the thickness of the glomerular membrane and basement membrane of proximal tubular epitheliocytes, as well as the width of podocyte foot processes and slit diaphragm were measured using scale bars.

Taskaeva I., Kasatova A., Surodin D., Bgatova N, & Taskaev S. (2023). Study of Lithium Biodistribution and Nephrotoxicity in Skin Melanoma Mice Model: The First Step towards Implementing Lithium Neutron Capture Therapy. Life, 13(2), 518.

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Publication 2023

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Histopathological Analysis of Rat Kidneys

Both kidneys were removed carefully after sacrificing rats without any damage and fixed with 10% neutral buffered formalin. Paraffin-embedded blocks were then made and sectioned (5 µm in thickness) for H&E, PAS, and Masson's trichrome staining to examine histopathological changes, glomerular basement membrane, and interstitial fibrosis, respectively (21 (link)). Images of stained kidney sections were taken using a Leica DM 2500 microscope (Leica Microsystems, Germany). For H&E-stained sections, ×400 magnification was used. For PAS- and Masson's trichrome-stained sections, ×1,000 magnifications were used. For each rat, separate slides were prepared. Ten different fields of the same slide were then examined. Two experienced renal pathologists assessed histopathological changes via quantitative tubulointerstitial injury measurement by counting numbers of apoptotic and necrotic cells, determining loss of tubular brush border, tubular dilatation, cast formation, and neutrophil infiltration, and examining glomeruli basement membrane thickness in a double-blinded fashion. The scoring was done based on level of damage: 0=none; 1=0-10%; 2=11-25%; 3=26-45%; 4=46-75%; and 5=76-100% (22 (link)).

Lee J.H., Islam M.S., Yoo Y.J., Kim S.E., Kim R.H., Jang Y.J., Lee S.H., Hwang H.P., Shin H.Y., Hwang J.H., Kim K., Park B.Y., Ahn D., Lee Y., Kim T., Kim I.S., Yoon J.C, & Tae H.J. (2023). Changes of antioxidant enzymes in the kidney after cardiac arrest in the rat model. Brazilian Journal of Medical and Biological Research, 56, e12408.

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Publication 2023

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Ultrastructural Analysis of Kidney after GOs Treatment

The ultrastructural changes of the kidney after GOs treatment were observed by TEM (JEOL JEM-1400) imaging. The kidney tissues were prefixed with 2.5% glutaraldehyde at 4 °C overnight and then post-fixed in 1% osmium tetroxide at 4 °C for 3 h. The samples were sectioned to 70 nm thick after dehydration and resin embedding, stained with uranyl acetate and lead citrate. The glomerular ultrastructural changes, including the glomerular basement membrane (GBM) thickness, podocyte foot process width and length, and glomerular slit diaphragm perimeter were quantitatively analyzed using ImageJ 1.51 k software.

Chen W., Wang B., Liang S., Wang M., Zheng L., Xu S., Wang J., Fang H., Yang P, & Feng W. (2023). Renal clearance of graphene oxide: glomerular filtration or tubular secretion and selective kidney injury association with its lateral dimension. Journal of Nanobiotechnology, 21, 51.

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Publication 2023

Citrate Dehydration Foot Glomerular Basement Membrane Glutaral Kidney Kidney Glomerulus Osmium Tetroxide Perimetry Podocytes Resins, Plant Tissues uranyl acetate Vaginal Diaphragm

Pathological Staging of Primary Membranous Nephropathy

By performing examinations on the patients, the following indicators are included: albumin (ALB), total cholesterol (TC), triglycerides (TG), blood urea nitrogen (BUN), creatinine (CREA), estimated glomerular filtration rate (eGFR), urine specific gravity (urine SG), urine red blood cells (urine RBC), and proteinuria. In addition, renal biopsy is performed on the patients, based on the results of renal biopsy, patients are divided into two groups: patients diagnosed with PMN and patients without any signs of PMN. For patients in the PMN group, the inclusion criteria are as follows: (1) patients are included if they are diagnosed with PMN clinically, (2) patients are diagnosed with PMN by examinations. For the control group, patients who are evaluated without evidence of PMN based on examinations and medical records are included. The characteristics of the patient data are summarized in Table 6.Table 6

Data statistics

Status	Value
Status	ALB	TC	TG	BUN	CREA	eGFR	Urine SG	Urine RBC (HPF)	Proteinuria
Non-PMN
Maximum	48.100	7.160	8.280	10.800	92.300	132.400	1.035	5.600	1.000
Minimum	30.900	2.750	0.440	2.900	43.100	86.500	1.004	0.100	0.000
Mean	43.772	4.362	1.561	5.431	68.847	118.270	0.955	1.156	0.172
PMN
Maximum	44.700	19.310	19.810	11.300	100.000	152.000	1.055	101.700	4.000
Minimum	12.400	4.270	0.510	2.200	30.000	78.000	1.002	0.300	0.500
Mean	26.056	8.514	2.768	4.817	65.510	108.873	1.019	13.496	2.490

Based on the appearance of electron-dense deposits in glomerular basement membrane (GBM) in electron microscopy, PMN can be classified into 4 stages. During the initial stages, podocyte effacement is noted with minimal to no changes in the GBM (stage I). If the deposits persist, new basement membrane material is laid between these immune deposits giving rise to the spike formations identified on methenamine silver stains which are readily observed on electron microscopy (stage II). In stage III, these deposits are completely encircled by newly laid basement membrane. In more advanced stages, basement membranes are thickened, and the deposits become more lucent and the spikes become less apparent. Our patients included only stage I and stage II, According to the renal biopsy results, among all patients with PMN, 61 patients were diagnosed with stage I and 41 with stage II, and no patients with pathological stage III–IV. Moreover, there are several non-PMN patients included in this study to further validate our approach. The distribution of patients are listed in Table 7.Table 7

Distribution of patients

	Number of patients
Non-PMN	32
Stage I PMN	61
Stage II PMN	41

Gao J., Wang S., Xu L., Wang J., Guo J., Wang H, & Sun J. (2023). Computer-aided diagnosis of primary membranous nephropathy using expert system. BioMedical Engineering OnLine, 22, 6.

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Publication 2023

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What is the function of the Glomerular Basement Membrane (GBM)?

The Glomerular Basement Membrane (GBM) is a specialized extracellular matrix structure that serves as the primary filtration barrier in the kidney's glomeruli. It is composed of collagen IV, laminin, and other components that provide structural support and selectively permit the passage of small molecules while retaining larger proteins. The GBM's key role is to facilitate the filtration of blood and the removal of waste products, while preventing the loss of important proteins.

What are the main components of the GBM?

How can PubCompare.ai assist in studying the GBM?

PubCompare.ai's AI-driven tools can help streamline your studies of the GBM by locating relevant protocols from the literature, preprints, and patents. The platform's comparison features can identify the most reproducible and accurate research methodologies, enabling you to choose the best approach for your specific research goals. This can save time and improve the efficacy of your GBM studies.

What are some common challenges in working with the GBM?

One of the main challenges in studying the GBM is understanding its complex composition and structure, as well as the intricate mechanisms involved in its filtration function. Additionally, accurately measuring and quantifying changes in the GBM's properties can be technichally dfficult. PubCompare.ai's tools can assist by helping you identify the most reliable and effective protocols to address these challenges.

Are there different types or variations of the GBM?

While the GBM is generally considered a single structure, there can be variations in its composition and ultrastructure depending on factors such as age, disease state, and location within the glomerulus. For example, the GBM may exhibit differences in its collagen IV and laminin isoforms, which can impact its filtration properties. Understanding these variations can be crucial for developing targeted therapies for GBM-related kidney diseases.

More about "Glomerular Basement Membrane"

The Glomerular Basement Membrane (GBM) is a critical component of the kidney's filtration system, serving as a specialized extracellular matrix that selectively permits the passage of small molecules while retaining larger proteins.
This vital structure is composed of collagen IV, laminin, and other essential elements that provide structural support and maintain the integrity of the glomeruli.
Alterations to the GBM's composition and ultrastructure have been linked to various kidney diseases, making it a crucial target for research into the underlying mechanisms and potential therapies.
Microscopic techniques, such as the JEM-1200EX II, H-7650, and JEM-1230 electron microscopes, as well as the Leica DM2500 light microscope and LSM 880 confocal laser scanning microscope, have been instrumental in studying the GBM's intricate structure and function.
The Embed 812 resin is commonly used for embedding and preparing samples for electron microscopy, while the use of Human IgG and Rabbit anti-human IgG antibodies can aid in the identification and visualization of specific proteins within the GBM.
By leveraging these advanced tools and techniques, researchers can gain valuable insights into the GBM's role in kidney health and disease, ultimately contributing to the development of more effective treatments.
PubCompare.ai's AI-driven optimization tools can streamline these studies by helping researchers locate relevant protocols from the literature, preprints, and patents, and identify the most reproducible and accurate research methodologies.
This can save time and resources, allowing researchers to focus on the most promising avenues of investigation and accelerate the understanding of the Glomerular Basement Membrane and its critical functions.