Abstract: SA-PO133
Whole-Kidney 3D Imaging for Assessment of Glomerular Number and Size in a Mouse Model of Diabetic Nephropathy
Session Information
- Diabetic Kidney Disease: Basic - III
October 27, 2018 | Location: Exhibit Hall, San Diego Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Diabetic Kidney Disease
- 601 Diabetic Kidney Disease: Basic
Authors
- Hecksher-Sørensen, Jacob, Gubra, Hørsholm, Denmark
- Roostalu, Urmas, Gubra, Hørsholm, Denmark
- Fabricius, Katrine, Gubra, Hørsholm, Denmark
- Vrang, Niels, Gubra, Hørsholm, Denmark
- Jelsing, Jacob, Gubra, Hørsholm, Denmark
- Secher, Thomas, Gubra, Hørsholm, Denmark
- Fink, Lisbeth N., Gubra, Hørsholm, Denmark
Background
Diabetic nephropathy (DN) is a major long-term complication of diabetes characterized by kidney hypertrophy and hyperfunction. To facilitate a rapid and unbiased evaluation of drug efficacy on glomerular size and number in preclinical studies, we investigated the use of light sheet microscopy as a new high-end 3D methodology to study whole-kidney glomerular changes in a mouse model of DN.
Methods
Unilateral nephrectomy (UNx) was performed in diabetic db/db mice to accelerate the development of nephropathy. UNx was performed in 18 weeks-old male db/db mice and terminated 6 weeks later. Mice were stratified and randomly assigned into UNx (n=8) or sham (n=8) based on baseline blood glucose levels. To determine the effect of UNx on whole-kidney glomerular morphology, mice were injected with lectin-594 prior to termination. Intact kidneys were scanned using light sheet microscopy (LSM). Using 3D image analysis, the total number of glomeruli were quantified and segmented according to individual size.
Results
Progression of diabetes remained similar in db/db control and db/db UNx mice. In contrast, terminal kidney weight was increased in db/db UNx mice, indicative of renal insufficiency leading to kidney hypertrophy. In agreement with stereology-based quantitative analyses on tissue sections, 3D-LSM confirmed an increase in kidney glomerular size in db/db UNx mice compared to db/db sham controls, while glomerular numbers were similar in both groups (≈14.000 glomeruli/kidney). Lectin-594 labelled kidneys were subsequently processed for Wilms' tumor-1 (Wt1), collagen IV and podocin expression using conventional immunohistochemistry. All antigens were readily detected, confirming that whole-kidney imaging was compatible for subsequent conventional immunohistochemistry.
Conclusion
In conclusion, LSM was successfully applied to evaluate renal and glomerular hypertrophy at the whole-organ level in a db/db UNx mice model of DN. Whole-kidney 3D imaging offers a novel approach for evaluating changes in key glomerular markers of DN, while maintaining the ability to perform conventional immunohistochemistry on the same tissue. The detailed analysis of all glomeruli enables for high-resolution evaluation of glomerular effects of novel treatment modalities in DN.