Stable expression of nephrin and localization to cell-cell contacts in novel murine podocyte cell lines

Schiwek, Daniel; Endlich, Nicole; Holzman, Lawrence B.; Holthöfer, Harry; Kriz, Wilhelm; Endlich, Karlhans

doi:10.1111/j.1523-1755.2004.00711.x

Cited by 123 publications

(139 citation statements)

References 45 publications

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“…94,95 Briefly, freshly isolated renal cortex was mixed and digested by collagenase I (2 mg/mL; Gibco, 17100-017) in RPMI 1640 (Life Technologies, 61870-044). Tissues were then passed through 70 mm and 40 mm cell strainers (BD falcon, 352350 and 352340).…”

Section: Assessment Of Renal Function and Albuminuriamentioning

confidence: 99%

Endothelial cell and podocyte autophagy synergistically protect from diabetes-induced glomerulosclerosis

et al. 2015

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filtration barrier; MAP1LC3A/B/LC3A/B), microtubule-associated protein 1 light chain 3 a/b; MTOR, mechanistic target of rapamycin; Nphs2, nephrosis 2, podocin; SQSTM1, sequestosome 1; STZ, streptozotocin; TEM, transmission electron microscopy; TUBA, tubulin; a, WT1, Wilms tumor 1.The glomerulus is a highly specialized capillary tuft, which under pressure filters large amounts of water and small solutes into the urinary space, while retaining albumin and large proteins. The glomerular filtration barrier (GFB) is a highly specialized filtration interface between blood and urine that is highly permeable to small and midsized solutes in plasma but relatively impermeable to macromolecules such as albumin. The integrity of the GFB is maintained by molecular interplay between its 3 layers: the glomerular endothelium, the glomerular basement membrane and podocytes, which are highly specialized postmitotic pericytes forming the outer part of the GFB. Abnormalities of glomerular ultrafiltration lead to the loss of proteins in urine and progressive renal insufficiency, underlining the importance of the GFB. Indeed, albuminuria is strongly predictive of the course of chronic nephropathies especially that of diabetic nephropathy (DN), a leading cause of renal insufficiency. We found that high glucose concentrations promote autophagy flux in podocyte cultures and that the abundance of LC3B II in podocytes is high in diabetic mice. Deletion of Atg5 specifically in podocytes resulted in accelerated diabetes-induced podocytopathy with a leaky GFB and glomerulosclerosis. Strikingly, genetic alteration of autophagy on the other side of the GFB involving the endothelialspecific deletion of Atg5 also resulted in capillary rarefaction and accelerated DN. Thus autophagy is a key protective mechanism on both cellular layers of the GFB suggesting autophagy as a promising new therapeutic strategy for DN.

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Section: Assessment Of Renal Function and Albuminuriamentioning

confidence: 99%

Endothelial cell and podocyte autophagy synergistically protect from diabetes-induced glomerulosclerosis

et al. 2015

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“…24 For differentiation, podocytes were cultured for 14 days under nonpermissive conditions (37°C, 7.4% CO 2 , RPMI 1640 supplemented with 10% fetal calf serum, 10 mmol/L N-2-hydroxyethylpiperazine-N0-2-ethanesulfonic acid, 1 mmol/L sodium pyruvate, 100 U/mL penicillin, and 100 mg/mL streptomycin). For proteasomal inhibition experiments, podocytes were cultured for 7 hours with 2 mol/L MG132 (Calbiochem, San Diego, CA) in DMSO or equal amounts of DMSO for control conditions.…”

Section: Cell Culturementioning

confidence: 99%

Ubiquitin C-Terminal Hydrolase-L1 Activity Induces Polyubiquitin Accumulation in Podocytes and Increases Proteinuria in Rat Membranous Nephropathy

Meyer-Schwesinger

Meyer

Sievert

et al. 2011

The American Journal of Pathology

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Ubiquitin C-terminal hydrolase L1 (UCH-L1), a key protease of the ubiquitin-proteasome system (UPS), is associated with neurodegenerative diseases and cancer. Recently, de novo expression of UCH-L1 was described in podocytes in patients with membranous nephropathy (MN), in which UCH-L1 expression correlated with increased ubiquitin content. The objective of the present study was to investigate the role of UCH-L1 in ubiquitin homeostasis and proteasomal degradation in a rat model of MN. After disease induction, UCH-L1 expression increased in podocytes and coincided with decreased glomerular monoubiquitin content. After an initial increase in proteasomal activity, the UPS was impaired. In addition to an increase of ubiquitin in podocytes, aggregates were observed 1 year after disease induction, as in MN in human beings. Inhibition of UCH-L1 hydrolase function in MN reduced UPS impairment and ameliorated proteinuria. In contrast, inhibition of proteasomal activity enhanced UPS impairment, resulting in increased proteinuria. Stable UCH-L1 overexpression in cultured podocytes resulted in accumulation of monoubiquitin and polyubiquitin proteins. In contrast, stable knockdown of UCH-L1 reduced monoubiquitin and polyubiquitin proteins and significantly increased proteasomal activity, indicating that the observed effects in rat MN also occurred in cultured podocytes. These data demonstrate that UCH-L1 activity results in polyubiquitin accumulation, proteasome inhibition, and disease aggravation in experimental models

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“…113 Briefly, podocytes were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium (PAA, Cölbe, Germany) supplemented with 10% fetal bovine serum (FBS) (Biochrom, Berlin, Germany) in a humidified atmosphere of 5% CO 2 . To stimulate podocyte proliferation, cells were cultivated at 33°C (permissive conditions).…”

Section: Cell Culturementioning

confidence: 99%

Mechanisms of angiotensin II signaling on cytoskeleton of podocytes

et al. 2008

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Stable expression of nephrin and localization to cell-cell contacts in novel murine podocyte cell lines

Cited by 123 publications

References 45 publications

Endothelial cell and podocyte autophagy synergistically protect from diabetes-induced glomerulosclerosis

Endothelial cell and podocyte autophagy synergistically protect from diabetes-induced glomerulosclerosis

Ubiquitin C-Terminal Hydrolase-L1 Activity Induces Polyubiquitin Accumulation in Podocytes and Increases Proteinuria in Rat Membranous Nephropathy

Mechanisms of angiotensin II signaling on cytoskeleton of podocytes

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