Polygonal epithelial cells in glomerular cell culture: Podocyte or parietal epithelial origin?

Yaoita, Eishin; Yoshida, Yutaka

doi:10.1002/jemt.10075

Cited by 9 publications

(8 citation statements)

References 37 publications

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“…Our findings are consistent with the observations of other investigators [3], [19], [21]–[23], who have already observed the distinct cellular morphologies. Interestingly, these investigators have also observed both morphologies emerging from glomeruli independent of the presence of a capsule with PECs.…”

Section: Discussionsupporting

confidence: 94%

Primary Cultures of Glomerular Parietal Epithelial Cells or Podocytes with Proven Origin

et al. 2012

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Parietal epithelial cells (PECs) are crucially involved in the pathogenesis of rapidly progressive glomerulonephritis (RPGN) as well as in focal and segmental glomerulosclerosis (FSGS). In this study, transgenic mouse lines were used to isolate pure, genetically tagged primary cultures of PECs or podocytes using FACsorting. By this approach, the morphology of primary glomerular epithelial cells in culture could be resolved: Primary podocytes formed either large cells with intracytoplasmatic extensions or smaller spindle shaped cells, depending on specific culture conditions. Primary PECs were small and exhibited a spindle-shaped or polygonal morphology. In the very early phases of primary culture, rapid changes in gene expression (e.g. of WT-1 and Pax-2) were observed. However, after prolonged culture primary PECs and podocytes still segregated clearly in a transcriptome analysis - demonstrating that the origin of primary cell cultures is important. Of the classical markers, synaptopodin and podoplanin expression were differentially regulated the most in primary PEC and podocyte cultures. However, no expression of any endogenous gene allowed to differentiate between the two cell types in culture. Finally, we show that the transcription factor WT1 is also expressed by PECs. In summary, genetic tagging of PECs and podocytes is a novel and necessary tool to derive pure primary cultures with proven origin. These cultures will be a powerful tool for the emerging field of parietal epithelial cell biology.

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Section: Discussionsupporting

confidence: 94%

Primary Cultures of Glomerular Parietal Epithelial Cells or Podocytes with Proven Origin

et al. 2012

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“…These findings corroborate the notion that they originate from PECs, which remains controversial. 57 GJIC has been demonstrated in vitro to have opposite effects on cell injury. Exposure of cells to irradiation or treatment with ganciclovir of herpes simplex virus thymidine kinase-transduced cells induces injury or death in not only the irradiated cells or the herpes simplex virus thymidine kinase-transduced cells but also adjacent nonirradiated cells or nontransduced cells.…”

Section: Discussionmentioning

confidence: 99%

Up-Regulation of Connexin43 in Glomerular Podocytes in Response to Injury

Yaoita

Yao

Yoshida

et al. 2002

The American Journal of Pathology

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Podocyte injury or podocyte loss in the renal glomerulus has been proposed as the crucial mechanism in the development of focal segmental glomerulosclerosis. However, it is poorly understood how podocytes respond to injury. In this study, glomerular expression of connexin43 (Cx43) gap junction protein was examined at both protein and transcript levels in an experimental model of podocyte injury, puromycin aminonucleoside (PAN) nephrosis. A striking increase in the number of immunoreactive dots with anti-Cx43 antibody was demonstrated along the glomerular capillary wall in the early to nephrotic stage of PAN nephrosis. The conspicuous change was not detected in the other areas including the mesangium and Bowman's capsule. Immunoelectron microscopy showed that the immunogold particles for Cx43 along the capillary wall were localized predominantly at the cell-cell contact sites of podocytes. Consistently, Western blotting and ribonuclease protection assay revealed a distinct increase of Cx43 protein, phosphorylation, and transcript in glomeruli during PAN nephrosis. The changes were detected by 6 hours after PAN injection. These findings indicate that the increase of Cx43 expression is one of the earliest responses that have ever been reported in podocyte injury. To show the presence of functional gap junctional intercellular communication (GJIC) in podocytes, GJIC was assessed in podocytes in the primary culture by transfer of fluorescent dye, Lucifer yellow, after a single-cell microinjection. Diffusion of the dye into adjacent cells was observed frequently in the cultured podocytes, but scarcely in cultured parietal epithelial cells of Bowman's capsule, which was compatible with their Cx43 staining. Thus, it is concluded that Cx43-mediated GJIC is present between podocytes, suggesting that podocytes may respond to injury as an integrated epithelium on a glomerulus rather than individually as a separate cell.

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“…Whereas it was previously reported that human glomerular epithelial cells were sensitive to Shiga toxin in vitro only at a much higher dose (27), the cells used prior were likely to be glomerular parietal epithelial cells rather than podocytes. This supposition is supported by their isolation using a sieving procedure shown to create cultures of nonpodocyte glomerular epithelial cells, their adoption of cobblestone as opposed to arborized morphology, and their lack of expression of the podocyte marker WT-1 (41,75,76).…”

Section: Vol 77 2009mentioning

confidence: 99%

Shiga Toxin 2 Targets the Murine Renal Collecting Duct Epithelium

et al. 2009

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Hemolytic-uremic syndrome (HUS) caused by Shiga toxin-producingEscherichia coli infection is a leading cause of pediatric acute renal failure. Bacterial toxins produced in the gut enter the circulation and cause a systemic toxemia and targeted cell damage. It had been previously shown that injection of Shiga toxin 2 (Stx2) and lipopolysaccharide (LPS) caused signs and symptoms of HUS in mice, but the mechanism leading to renal failure remained uncharacterized. The current study elucidated that murine cells of the glomerular filtration barrier were unresponsive to Stx2 because they lacked the receptor glycosphingolipid globotriaosylceramide (Gb 3 ) in vitro and in vivo. In contrast to the analogous human cells, Stx2 did not alter inflammatory kinase activity, cytokine release, or cell viability of the murine glomerular cells. However, murine renal cortical and medullary tubular cells expressed Gb 3 and responded to Stx2 by undergoing apoptosis. Stx2-induced loss of functioning collecting ducts in vivo caused production of increased dilute urine, resulted in dehydration, and contributed to renal failure. Stx2-mediated renal dysfunction was ameliorated by administration of the nonselective caspase inhibitor Q-VD-OPH in vivo. Stx2 therefore targets the murine collecting duct, and this Stx2-induced injury can be blocked by inhibitors of apoptosis in vivo.

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Polygonal epithelial cells in glomerular cell culture: Podocyte or parietal epithelial origin?

Cited by 9 publications

References 37 publications

Primary Cultures of Glomerular Parietal Epithelial Cells or Podocytes with Proven Origin

Primary Cultures of Glomerular Parietal Epithelial Cells or Podocytes with Proven Origin

Up-Regulation of Connexin43 in Glomerular Podocytes in Response to Injury

Shiga Toxin 2 Targets the Murine Renal Collecting Duct Epithelium

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