Abstract
Given that podocalyxin (PCX) and nestin play important roles in podocyte morphogenesis and the maintenance of structural integrity, we examined whether the expression and localization of these two podocyte proteins were influenced in the early stage of various hemodynamic conditions. Mice kidney tissues were prepared by in vivo cryotechnique (IVCT). The distribution of glomeruli and podocyte proteins was visualized with DAB staining, confocal laser scanning microscopy and immunoelectron microscopy. The mRNA levels were examined by real-time quantitative PCR. The results showed the following: Under the normal condition, PCX stained intensely along glomerular epithelial cells, whereas nestin was clearly staining in the endothelial cells and appeared only weakly in the podocytes. Under the acute hypertensive and cardiac arrest conditions, PCX and nestin staining was not clear, with a disarranged distribution, but the colocalization of PCX and nestin was apparent under this condition. In addition, under the acute hypertensive and cardiac arrest conditions, the mRNA levels of PCX and nestin were significantly decreased. Collectively, the abnormal redistribution and decreased mRNA expressions of PCX and nestin are important molecular events at the early stage of podocyte injury during hemodynamic disorders. IVCT may have more advantages for morphological analysis when researching renal diseases.
Highlights
The glomerular podocyte is a terminally differentiated cell that lines the outer aspect of the glomerular basement membrane (GBM)
For the sections obtained by in vivo cryotechnique (IVCT), the capillary loops were smooth under a normotensive condition (Figure 1A); under the acute hypertensive condition, the Bowman’s space and luminal spaces of the proximal and distal tubules were open widely in the renal cortices (Figure 1B), and neither the Bowman’s spaces nor luminal spaces of the tubules were clearly open under the cardiac arrest condition (Figure 1C)
These results indicate that the acute hypertensive and cardiac arrest conditions decrease the expression of PCX and nestin and influence the localization of these podocyte proteins
Summary
The glomerular podocyte is a terminally differentiated cell that lines the outer aspect of the glomerular basement membrane (GBM). Nestin is an intermediate filament protein originally described in neural stem cells and is recently thought to be expressed in differentiated podocytes of the adult kidney, which might be associated with the maintenance of the foot process structure [11,12]. Based on the functions and possible linkage of PCX and nestin, we examined the changes in their expression and localization under various hemodynamic conditions, especially during the early stage, which is still poorly understood. Hemodynamic factors, such as blood flow and pressure, are well-known to exert an important influence on the kidney structure and function, and these effects occur almost instantaneously [16]. Our findings provide a new insight into the early molecular mechanisms of podocyte injury during hemodynamic disorders
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