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Pathobiology. 1996;64(4):222-32.

Biological alterations of rat podocytes cultured under basolateral hydrostatic pressure.

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  • 1Department of Pathology, University of Groningen, The Netherlands.

Abstract

In vivo, glomerular visceral epithelial cells (GVEC), or podocytes, are morphologically highly differentiated cells which are in close contact with adjacent cells by complex interdigitating foot processes. In vitro, the dedifferentiated appearance of podocytes hampers investigations on podocyte structure and function. Cultured podocytes resemble simple epithelium in several ways with apical tight junctions and absence of foot processes. The morphological resemblances between GVEC early in embryonic development, in proteinuric diseases and in cultured cells are striking, but the mechanisms involved in these (de)differentiation processes are poorly understood. A common feature of GVEC in these various states of dedifferentiation is their altered exposure to or even total lack of hydrostatic pressure, suggesting that this may be one of the parameters involved in GVEC differentiation. In this study we investigated whether basolateral hydrostatic pressure could affect GVEC biology in vitro. We therefore exposed cultured GVEC grown on porous supports to basolateral hydrostatic pressure and investigated morphology with scanning and transmission electron microscopy, expression of specific podocyte markers and their biological responses to a model stimulus, the cytokine IFN-gamma. Morphologically, monolayers of pressurized GVEC contained large regions of whirl-like, raised cell formations. Individual cells in these formations had a rounded morphology and pore-like indentations between adjacent cells were observed. Cell-cell contacts were often found more basally and intercellular spaces were widened. Moreover, protein expression of pressurized monolayers was altered, as demonstrated by regions of cells with decreased keratin expression. Finally, upon exposure to the model stimulus IFN-gamma, the pressurized as compared to the control GVEC demonstrated a 3-fold increased expression of MHC class II and a strongly decreased sensitivity to the toxic effects of IFN-gamma. In conclusion, we found several indications that hydrostatic pressure can affect podocyte biology in vitro and similar mechanisms may account for podocyte biology in vivo. The strikingly altered morphology and biology of pressurized GVEC suggest that this culture system can be quite relevant for future studies with cultured GVEC.

PMID:
9031333
[PubMed - indexed for MEDLINE]
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