Using Stereologic Techniques for Podocyte Counting in the Mouse: Shifting the Paradigm
Open Access
- 1 June 2011
- journal article
- research article
- Published by S. Karger AG in American Journal of Nephrology
- Vol. 33 (Suppl. 1), 1-7
- https://doi.org/10.1159/000327564
Abstract
Background: The podocyte serves the important function of maintaining the glomerular filtration barrier, and many studies report a decrease in podocyte number relative to the development of proteinuric states. However, there is significant inconsistency in the number of podocytes counted, possibly due to different counting methods. We previously counted the three glomerular cell types in the mouse kidney and showed that the fractionator/disector method is a close approximation of the exhaustive count or the gold standard method. In this study, we compared the commonly used model-based approach with the design-based approach to count podocytes in the db/m and db/db mouse and illustrate that the design-based approach, which uses the fractionator/disector method, provides an accurate determination of podocyte number. Methods: In the design-based approach, toluidine blue-stained 1-µm-thick sections from glutaraldehyde perfusion-fixed kidneys were used (n = 15) with the fractionator/disector method. In the model-based approach, WT-1-immunolabeled podocyte nuclei in 3- to 4-µm-thick formalin-fixed paraffin-embedded sections of the same kidneys were counted with the Weibel-Gomez method. Glomerular volume was determined for each method. Results: We discovered that the fractionator/disector method counted 89 ± 10 podocytes compared to the Weibel-Gomez method, which counted 137 ± 38 podocytes and overestimated podocyte number by 54% (p < 0.05). In addition, glomerular volume (231 ± 52 × 103 vs. 192 ± 64 × 103 µm3) was significantly underestimated by 17% (p < 0.0002). Moreover, the model-based approach was more time consuming. Conclusion: We conclude that the fractionator/disector method offers an unbiased and efficient determination of podocyte counts.Keywords
This publication has 27 references indexed in Scilit:
- Podocyte Number in the Maturing Rat KidneyAmerican Journal of Nephrology, 2010
- Facebook activity of residents and fellows and its impact on the doctor-patient relationshipJournal of Medical Ethics, 2010
- Online Social Networking by Patients with Diabetes: A Qualitative Evaluation of Communication with FacebookJournal of General Internal Medicine, 2010
- Targeting podocyte-associated diseases☆Advanced Drug Delivery Reviews, 2010
- Osteopontin modulates angiotensin II–induced inflammation, oxidative stress, and fibrosis of the kidneyKidney International, 2009
- Recruitment of Podocytes from Glomerular Parietal Epithelial CellsJournal of the American Society of Nephrology, 2009
- Podocyte Loss in Aging OVE26 Diabetic MiceThe Anatomical Record, 2007
- Comparison of Methods for Counting Cells in the Mouse GlomerulusNephron Experimental Nephrology, 2006
- Estimation of podocyte number: A comparison of methodsKidney International, 2004
- Wilkins, Crick, and Watson: Nobel Prize for Work on the Structure of DNAMayo Clinic Proceedings, 1998