Recent evidence suggests that glomerular hypertrophy is a key event in the development of focal and segmental glomerulosclerosis and hyalinosis (FSGS) in humans and in many experimental models of FSGS. The initial aim of the present study was to determine if glomerular hypertrophy occurs in a puromycin aminonucleoside (PAN) model of FSGS, previously considered not to involve glomerular hypertrophy. Upon identifying significant glomerular hypertrophy, our second aim was to determine the contribution of glomerular capillary growth to this hypertrophy. Female Sprague-Dawley rats (approximately 200 g) were administered either PAN (2 mg/100 g body wt) subcutaneously, or an equivalent volume of 0.9% saline at weeks 0, 1, 2, 4, 6, 8 and 10. Tissue was analyzed at weeks 7 and 13. Unbiased stereological methods were used to estimate a range of glomerular parameters. Mean glomerular tuft volume in PAN-treated rats was 48% greater than in saline-treated rats at seven weeks, and 63% greater at 13 weeks. Similar results were found for mean renal corpuscle volume. FSGS was absent at seven weeks and minor at 13 weeks. Two-way analysis of variance indicated: significant effects (P < 0.05 at least) of PAN on capillary length per glomerulus, capillary surface area per glomerulus, capillary diameter and length of capillaries per unit volume of glomerulus; and significant effects of time on capillary diameter, capillary length per unit volume of glomerulus and capillary surface area per unit volume of glomerulus. The mean length of capillaries per glomerulus was 45% greater in PAN-treated rats at week 7 and 22% greater in PAN-treated rats at week 13. Taken together, these results indicate a biphasic pattern of glomerular hypertrophy in this model. In the first phase (to 7 weeks), an increase in capillary length contributes to glomerular hypertrophy. In the second phase (7 to 13 weeks), the continued glomerular enlargement appears more likely to be due to an increase in capillary diameter and/or mesangial matrix expansion.

Summary:Quantitative methods are frequently used to analyse the structure of renal glomeruli. However, on most occasions, measurements are made on glomerular profiles (the two-dimensional samples of glomeruli seen in histological sections), and provide little or no information about the structure of whole, three-dimensional glomeruli. Stereology is the discipline concerned with the quantitative analysis of three-dimensional structures. With stereology one can estimate the total number of glomeruli in kidneys, as well as mean glomerular volume, the number of cells in glomeruli, and the length and surface area of glomerular capillaries. In addition to providing a means for detecting structural differences between glomeruli from different specimens, stereology provides quantitative structural information that can be correlated with quantitative physiological, biochemical and molecular data. Over the past decade we have witnessed the development of a new generation of unbiased, cost-efficient stereological methods that are ideally suited to analysing glomeruli. Some of these methods are introduced in this review, and then three recent studies from our laboratories that successfully utilized these methods are described. These studies concerned hypertension, kidney development, and the pathogenesis of focal and segmental glomerulosclerosis.

Focal and segmental glomerulosclerosis (FSGS) is a chronic renal disorder characterized by segmental glomerular lesions and widespread podocyte foot process effacement. We have previously shown that glomerular enlargement (hypertrophy) precedes the development of FSGS in an animal model not previously thought to involve glomerular hypertrophy. This hypertrophy involved growth of glomerular capillaries. The aim of the present study was to determine whether the capillary growth involved an increase in the number of capillaries per glomerulus, or lengthening of existing capillaries. In addition, we examined the contribution of glomerular cell hyperplasia to the hypertrophy. We found that glomerular capillary growth in this model appears to primarily involve lengthening of existing capillaries rather that sprouting of new capillaries, and that glomerular cell proliferation contributes to the glomerular hypertrophy.

Focal and segmental glomerulosclerosis (FSGS) is a chronic renal disorder characterized by segmental glomerular lesions and widespread podocyte foot process effacement. We have previously shown that glomerular enlargement (hypertrophy) precedes the development of FSGS in an animal model not previously thought to involve glomerular hypertrophy. This hypertrophy involved growth of glomerular capillaries. The aim of the present study was to determine whether the capillary growth involved an increase in the number of capillaries per glomerulus, or lengthening of existing capillaries. In addition, we examined the contribution of glomerular cell hyperplasia to the hypertrophy. We found that glomerular capillary growth in this model appears to primarily involve lengthening of existing capillaries rather that sprouting of new capillaries, and that glomerular cell proliferation contributes to the glomerular hypertrophy.