Measuring the Volume Density of the Glomerular Mesangium

Jm, Basgen; Rich, Stephen S.; Sm, Mauer; Mw, Steffes

doi:10.1159/000185154

Cited by 15 publications

(3 citation statements)

References 9 publications

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“…Measurements of relative mesangial per glomerular volume (Vv(mes/glom) was performed using the point grid system and Image Pro Plus image analysis software as described previously 17–19 . In brief, ultra-thin renal sections (70 – 80 nm) were obtained and examined with a Philips 410LS Transmission Electron Microscope in the Dept.…”

Section: Methodsmentioning

confidence: 99%

Simvastatin Reverses Podocyte Injury but not Mesangial Expansion in Early Stage Type 2 Diabetes Mellitus

et al. 2009

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Statins may confer renal protection in a variety of glomerular diseases, including diabetic nephropathy (DN). However, various glomerular lesions have different etiologies and may have different responses to statins. This study was performed to determine the differential effects of simvastatin (SMV) on glomerular pathology including mesangial expansion and podocyte injury in a mouse model of early stage type 2 diabetes mellitus (DM). Type 2 DM was induced in male C57BL/6 mice by feeding a high fat diet (HF; 45 kcal% fat). After 22 weeks, one group of HF mice was treated with SMV (HF-SMV; 7 μg/day/g BW) and another group was treated with vehicle (HF-vehicle) for 4 weeks via osmotic mini-pump. A third group served as age-matched normal diet vehicle controls (ND-vehicle; 10 kcal% fat). At the end of treatment, glomerular morphology was evaluated in a blind manner to determine the progression of DN. Body weight, blood glucose, insulin, HDL-cholesterol and triglycerides, but not LDL-cholesterol, were increased in HF mice. Over the course of treatment, the 24-hour urinary albumin excretion (UAE) was unchanged in ND-vehicle. HF mice exhibited elevated UAE, which decreased with SMV, but was unchanged with vehicle. The absolute mesangial volume and the relative mesangial volume per glomerular volume increased in HF-vehicle and remained elevated with SMV treatment. The immuno-staining of nephrin, a protein marker of the integrity of podocyte slit diaphragms, was decreased in HF-vehicle; however, the nephrin quantity of the HF-SMV group was not different from ND-vehicle. It is concluded that SMV reverses podocyte damage, but does not affect mesangial expansion in the kidneys of early stage proteinuria of type 2 DM.

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Section: Methodsmentioning

confidence: 99%

Simvastatin Reverses Podocyte Injury but not Mesangial Expansion in Early Stage Type 2 Diabetes Mellitus

et al. 2009

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“…6A). To determine the minimum number of image samples required to measure accurately the model and real mesangial volume fraction, we reviewed the point-counting data of Basgen et al (1988). In this previous study, human glomerular mesangial expansion (67 percent increase above normal) was detected in early diabetic glomerulosclerosis.…”

Section: Methodsmentioning

confidence: 99%

A new computer‐assisted three‐dimensional reconstruction method provides accurate measurement of glomerular mesangial volume

Cho

Winslow

Whiteside

et al. 1991

J. Elec. Microsc. Tech.

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Many glomerulopathies are characterized by progressive mesangial (interstitial) expansion which can be quantitated by morphometric analysis. The purpose of this study was to analyze mesangial and glomerular volumes using a new computer-assisted reconstruction (CAR) method. CAR was compared to two standard planar methods, point-counting and linear integration, for accuracy and time efficiency. In Phase I of the study, a computer-based model of the mesangial space was created by placing spherical and ellipsoidal objects of known volume into an enclosing volume mimicking the glomerulus. The simulated mesangium occupied approximately 10 percent of the glomerular volume. The model glomerulus was sectioned serially into ten sections of equal thickness and the three morphometric methods applied to determine the mesangial/glomerular volume. The complexity of the mesangial model was varied by increasing the number of mesangial regions from one to ten to 100. The CAR method estimated the model mesangial volume more accurately (1-9 percent error) through each level of complexity compared to point-counting (3-17 percent error) and linear integration (3-18 percent error). The point-counting method consistently overestimated (P less than 0.05) the fractional mesangial volume for the ten- and 100-region mesangium models. In Phase II of the study, a normal rat glomerulus was sectioned serially (215 sections) and a transmission electron micrograph (TEM) of every fifth section (n = 43) was obtained. Each TEM image (2% of glomerular surface) was digitized for analysis by CAR. Point-counting and linear integration were also performed on the whole glomerular TEMs (n = 10, randomly chosen). The estimated relative mesangial/glomerular volume was 6.6 +/- 0.1 percent by CAR (mean +/- SD), 9.7 +/- 1.5 by linear integration, and 14.9 +/- 3.4 by point-counting. The point-counting method was most efficient, requiring 40 +/- 8 sec/section, followed by CAR at 85 +/- 24 sec/section. Linear integration was least efficient (93 +/- 23 sec/section). We conclude that CAR is the most accurate morphometric method of the three compared for estimating mesangial and glomerular methods, although it is more time consuming than the point-counting method and requires more complex instrumentation. CAR is the only method that will analyze the shape and three-dimensional complexity of glomerular structures using TEMs.

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“…These Image enhancement and tracing techniques can be quite time consuming. Often a more efficient measuring method is to superimpose an appropriate counting grid of points and/or lines over the image [14]. Grid points falling on profiles are counted to estimate areas, intercepts between grid lines and component lines are counted to estimate lengths, and number of component profiles is counted when determining length and number density.…”

Section: Number Densitymentioning

confidence: 99%

Basic Stereology

Basgen¹

2003

Microscopy Today

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Many us who use microscopes are interested in the internal structure or components of three-dimensional objects. Often we must section these objects to observe these internal components. For many years, microtomes have been used to make physical sections, but in recent years confocal microscopes, MR imaging, CT scanners, and even standard optical microscopes have been used to obtain “optical” sections. Two-dimensional images of these different types of sections can be used to extract three-dimensional quantitative information about the objects and their internal components, The sectioning process reduces the observed dimensions of the object and components. With apologies to Rene Magritte, the structure portrayed in Figure 1 is not a three-dimensional glomerulus but a two-dimensional profile of a glomerulus. In most cases, interest is on the structure of the three-dimensional object and not the structure in the two-dimensional image. Thus, care must be taken when obtaining and interpreting data from two-dimensional images.

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Measuring the Volume Density of the Glomerular Mesangium

Cited by 15 publications

References 9 publications

Simvastatin Reverses Podocyte Injury but not Mesangial Expansion in Early Stage Type 2 Diabetes Mellitus

Simvastatin Reverses Podocyte Injury but not Mesangial Expansion in Early Stage Type 2 Diabetes Mellitus

A new computer‐assisted three‐dimensional reconstruction method provides accurate measurement of glomerular mesangial volume

Basic Stereology

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