Abstract
Rarefaction of the renal microvasculature correlates with declining kidney function. However, current technologies commonly used for its evaluation are limited by their reliance on endothelial cell antigen expression and assessment in two dimensions. We set out to establish a widely applicable and unbiased optical sectioning method to enable three dimensional imaging and reconstruction of the renal microvessels based on their luminal filling. The kidneys of subtotally nephrectomized (SNx) rats and their sham-operated counterparts were subjected to either routine two-dimensional immunohistochemistry or the novel technique of fluorescent microangiography (FMA). The latter was achieved by perfusion of the kidney with an agarose suspension of fluorescent polystyrene microspheres followed by optical sectioning of 200 µm thick cross-sections using a confocal microscope. The fluorescent microangiography method enabled the three-dimensional reconstruction of virtual microvascular casts and confirmed a reduction in both glomerular and peritubular capillary density in the kidneys of SNx rats, despite an overall increase in glomerular volume. FMA is an uncomplicated technique for evaluating the renal microvasculature that circumvents many of the limitations imposed by conventional analysis of two-dimensional tissue sections.
Highlights
Glomerular and peritubular capillary loss are major histopathological features of most forms of chronic kidney disease (CKD) that, by predisposing to tissue hypoxia, are fundamentally linked to its progression [1]
In comparison to sham-operated animals, SNx rats demonstrated an increase in systolic blood pressure (SBP) and urine protein excretion with a decline in glomerular filtration rate (GFR) (Table 1)
Assessment of the renal vasculature has been achieved using a range of antibodies directed against endothelial cell surface antigens
Summary
Glomerular and peritubular capillary loss are major histopathological features of most forms of chronic kidney disease (CKD) that, by predisposing to tissue hypoxia, are fundamentally linked to its progression [1]. Laboratory methods for the detailed assessment of the kidney microvasculature are overly labor intensive and the vast majority of studies exploring the therapeutic efficacy of novel agents rely on conventional light microscopic examination of antibody- or lectin-stained kidney sections. Such techniques, while simple to perform, are clearly limited by their assessment in two dimensions [3], and by the lack of specificity of the labeling techniques employed [4]. The utility of this approach for assessing the microvascular architecture of the kidney in the physiological and disease settings was assessed in sham-operated and subtotally nephrectomized (SNx) rats, respectively
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