Words similar to podocytes
Example sentences for: podocytes
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Protein (250 μg) from the control-infected or HIV-1-infected podocytes on day 14 of the differentiation protocol was also analyzed by western blot for total pRb (ECL detection of a 1:1000 dilution of rabbit anti-Rb, Santa Cruz Biotechnology) and phospho-Rb (Ser780) (ECL detection of a 1:500 dilution of rabbit anti-phospho-Rb (Ser780) antibody binding, Cell Signaling).
These data indicate that HIV-1 expression in infected podocytes induces cyclin D 1 and phospho-pRb (Ser780) expression, hallmarks of cyclin D 1 -mediated G 1 → S progression.
Here, we ask whether infected podocytes expressing HIV-1 genes show the hallmarks of cyclin D 1 -dependent proliferation, specifically the up-regulation of cyclin D 1 transcript and protein expression and the subsequent phosphorylation of pRb on serine 780, a target site of active cyclin D 1 /CDK-4/6 that is not shared with cyclin E/CDK-2 complexes [ 24 ] . Utilizing two well-characterized models of HIVAN pathogenesis, HIV-1 transgenic mice [ 11 12 13 ] and HIV-1 infection of cultured podocytes [ 6 7 8 ] we find not only these hallmarks of cyclin D 1 -dependent proliferation, but also dysregulation of cyclin D 1 expression that normally occurs with cell-cell contact and differentiation [ 25 ] .
In order to determine if cyclin D 1 plays a role in this hyperplasia, kidneys from three proteinuric 40-day old transgenic siblings and three non-transgenic littermates were stained by immunohistochemistry with antibody to phospho-pRb (Ser780), a site of cyclin D 1 /CDK-4/6 phosphorylation not targeted by cyclin E/CDK-2 complexes that disrupts pRb/E2F binding for G 1 → S progression [ 24 ] . Figure 1depicts the intense, heterogeneous nuclear staining of glomerular podocytes and tubular epithelium from hyperplastic nephrons in the diseased transgenics versus the absent staining of nephrons in the non-transgenics consistent with the very low mitotic index in normal kidneys of adult animals [ 26 ] . This suggests that cyclin D 1 is promoting cell-cycle progression during the proliferation of epithelium in diseased kidneys.
The ongoing proliferation of HIV-1-infected podocytes following cell-cell contact led to the formation of multiple foci (Figure 2C) with intense, heterogenous nuclear cyclin D 1 immunofluorescence consistent with ongoing asynchronous proliferation in this population of cells (Figure 3).
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