Three-Dimensional Morphology by Multiphoton Microscopy with Clearing in a Model of Cisplatin-Induced CKD

Abstract

Traditional histologic methods are limited in their ability to detect pathologic changes of CKD, of which cisplatin therapy is an important cause. In addition, poor reproducibility of available methods has limited analysis of the role of fibrosis in CKD. Highly labor-intensive serial sectioning studies have demonstrated that three-dimensional perspective can reveal useful morphologic information on cisplatin-induced CKD. By applying the new technique of multiphoton microscopy (MPM) with clearing to a new mouse model of cisplatin-induced CKD, we obtained detailed morphologic and collagen reconstructions of millimeterthick renal sections that provided new insights into pathophysiology. Quantitative analysis revealed that a major long-term cisplatin effect is reduction in the number of cuboidal cells of the glomerular capsule, a change we term the "uncapped glomerulus lesion." Glomerulotubular disconnection was confirmed, but connection remnants between damaged tubules and atubular glomeruli were observed. Reductions in normal glomerular capsules corresponded to reductions in GFR. Mild increases in collagen were noted, but the fibrosis was not spatially correlated with atubular glomeruli. Glomerular volume and number remained unaltered with cisplatin exposure, but cortical tubulointerstitial mass decreased. In conclusion, new observations were made possible by using clearing MPM, demonstrating the utility of this technique for studies of renal disease. This technique should prove valuable for further characterizing the evolution of CKD with cisplatin therapy and of other conditions.

Non-Transgenic Mouse Models of Kidney Disease

“…Torres et al [12] developed a murine model of CKD based on repetitive cisplatin administrations. Two doses of i.p.…”

“…Kidneys decreased in volume and showed myofibroblast proliferation, increased apoptosis and loss of interglomerular mass by light microscopy and immunohistochemistry. Three-dimensional microscopy revealed a reduction of cuboidal cells of the glomerular capsule and glomerulotubular disconnection [12]. …”

Age-Related Renal Microvascular Changes: Evaluation by Three-Dimensional Digital Imaging of the Human Renal Microcirculation Using Virtual Microscopy

“…Recent technical advances have enabled us to reconstruct high-resolution 3D images of the renal microvasculature, which have proved useful for investigating renal damage caused by an altered microvasculature [21,22,23,24,25]. Micro-computed tomography with contrast media can successfully quantify vessel density and vascular volume and has confirmed the rarefaction of renal capillaries in eNOS −/− rat kidney [21].…”

“…Confocal microscopy with fluorescence micro-angiography proved the reduction of glomerular and peritubular capillary density [1] and the increase in capillary branching of glomeruli [22,23] in a diseased rat kidney. Recently, Torres et al created 3D images of the whole kidney in cisplatin-treated mice and successfully reconstructed a complete nephron using multi-photon microscopy [24]. …”

A practical new way to measure kidney fibrosis

“…This would provide additional detail on focal fibrotic patterns, as was shown recently. 8 Also, future studies will likely apply and test these techniques for live animal imaging. Tracking the development and progression of the fibrosis process in the same animal and tissue region over time, as shown recently for tracking individual cell types, 9 would further expand the capabilities of this SHG-FLIM—based approach.…”