Total numbers of glomeruli and individual glomerular cell types in the normal rat kidney

Alterations in numbers of glomeruli and glomerular cells occur in various renal disorders. Although values for these parameters have previously been reported for several species, the estimates have often been biased due to assumptions regarding glomerular and/or nuclear size and shape. Other studies have used tedious serial-section reconstruction methods. In the present study, unbiased stereological methods were used to estimate total numbers of glomeruli and individual glomerular cell types in normal rats. The kidneys of seven adult Sprague-Dawley rats were perfused with 4% paraformaldehyde and 1% glutaraldehyde in phosphate buffer and embedded in either glycol-methacrylate (for light microscopy, LM) or Epon/Araldite (for transmission electron microscopy, TEM). Total glomerular number was estimated using an LM physical disector/fractionator combination; the total number of cells per average glomerulus was estimated using an LM optical disector/Cavalieri combination; and TEM physical disectors were used to count individual cell types. The normal rat kidney was found to contain 31,764 +/- 3667 (mean +/- SD) glomeruli. An average glomerulus contained 674 +/- 129 cells, of which 181 +/- 53 were epithelial cells (podocytes), 248 +/- 53 were endothelial cells, and 245 +/- 45 were mesangial cells. An average renal corpuscle contained 117 +/- 27 parietal epithelial cells. Following sectioning and staining, less than 6.5 h was needed to obtain the above estimates for a single animal, with coefficients of variation (SD as a percent of the mean) ranging from 10% to 25%. The unbiased stereological methods used in the present study constitute an unbiased, precise and cost-efficient set of quantitative tools for assessing glomerular morphology in health and disease.