Browse
Submit Data
Databases
API
Help

Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

11 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Mechanical stress enhances CD9 expression in cultured podocytes

ABSTRACT: Transcriptomes of differentiated cells of the conditionally immortalized mouse podocyte cell line SVI (Schiwek et al., Kidney Int. 66: 91-101, 2004) were determined as described in Warsow et al. (Kidney Int. 84: 104-115, 2013) after application of mechanical stress (Endlich et al., J. Am. Soc. Nephrol. 12: 413-422, 2001) as compared to control conditions. Elevated glomerular pressure represents a high risk for the development of severe kidney diseases and causes an increase of mechanical load to podocytes. In this study we investigated whether mechanical stress alters gene expression in cultured podocytes using gene arrays. We found that tetraspanin CD9 is significantly upregulated in cultured podocytes after mechanical stress. The differential expression of CD9 was confirmed by RT-PCR and Western blot under stretched and unstretched conditions. Furthermore, mechanical stress resulted in a relocalization of CD9. To get an insight into the functional role of CD9, podocytes were transfected with pEGFP-CD9. The expression of CD9 induced the formation of substratum-attached thin arborized protrusions (TAPs). Ca2+ depletion revealed that podocytes over-expressing CD9 possess altered adhesive properties in contrast to the control transfected cells. Finally, elevated CD9 expression increased migration of podocytes in a wound assay. In summary, our results suggest that upregulation of CD9 may play an important role in podocyte morphology, adhesion and migration. Three independent batches were used.

ORGANISM(S): Mus musculus

SUBMITTER: Karlhans Endlich

PROVIDER: E-GEOD-66336 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Similar Datasets

Mechanical stress enhances CD9 expression in cultured podocytes

Project description:Transcriptomes of differentiated cells of the conditionally immortalized mouse podocyte cell line SVI (Schiwek et al., Kidney Int. 66: 91-101, 2004) were determined as described in Warsow et al. (Kidney Int. 84: 104-115, 2013) after application of mechanical stress (Endlich et al., J. Am. Soc. Nephrol. 12: 413-422, 2001) as compared to control conditions. Elevated glomerular pressure represents a high risk for the development of severe kidney diseases and causes an increase of mechanical load to podocytes. In this study we investigated whether mechanical stress alters gene expression in cultured podocytes using gene arrays. We found that tetraspanin CD9 is significantly upregulated in cultured podocytes after mechanical stress. The differential expression of CD9 was confirmed by RT-PCR and Western blot under stretched and unstretched conditions. Furthermore, mechanical stress resulted in a relocalization of CD9. To get an insight into the functional role of CD9, podocytes were transfected with pEGFP-CD9. The expression of CD9 induced the formation of substratum-attached thin arborized protrusions (TAPs). Ca2+ depletion revealed that podocytes over-expressing CD9 possess altered adhesive properties in contrast to the control transfected cells. Finally, elevated CD9 expression increased migration of podocytes in a wound assay. In summary, our results suggest that upregulation of CD9 may play an important role in podocyte morphology, adhesion and migration.

2015-02-27 | GSE66336 | GEO

Differentiated mouse podocytes (SVI)

Project description:Transcriptomes of differentiated cells of the conditionally immortalized mouse podocyte cell line SVI (Schiwek et al., Kidney Int. 66: 91-101, 2004) were determined as described in Kabgani et al. (PLoS One 7:e34907, 2012). The transcriptomes of the podocyte cell line were mapped on a protein-protein interaction network of the podocyte (PodNet). Together with other transcriptomes taken from GEO, we analyzed differential gene regulation and differential regulation of protein-protein interactions between cultured podocytes and differentiated in vivo podocytes.

2012-10-16 | GSE41523 | GEO

Differentiated mouse podocytes (SVI)

Project description:Transcriptomes of differentiated cells of the conditionally immortalized mouse podocyte cell line SVI (Schiwek et al., Kidney Int. 66: 91-101, 2004) were determined as described in Kabgani et al. (PLoS One 7:e34907, 2012). The transcriptomes of the podocyte cell line were mapped on a protein-protein interaction network of the podocyte (PodNet). Together with other transcriptomes taken from GEO, we analyzed differential gene regulation and differential regulation of protein-protein interactions between cultured podocytes and differentiated in vivo podocytes. Three independent batches were used.

2012-10-16 | E-GEOD-41523 | biostudies-arrayexpress

Mus musculus

Project description:Mechanical stress enhances CD9 expression in cultured podocytes

| PRJNA276570 | ENA

Podocyte-specific deletion of tubular sclerosis complex 2 causes focal segmental glomerular sclerosis and progressive renal failure

Project description:Obesity can initiate and accelerate the progression of kidney diseases. However, it remains unclear how obesity affects renal dysfunction. Here, we show that a newly generated podocyte-specific tubular sclerosis complex 2 (Tsc2) knockout mouse model (Tsc2∆podocyte) develops proteinuria and dies due to end-stage renal dysfunction by 10 weeks of age. Tsc2∆podocyte mice exhibit an increased glomerular size and focal segmental glomerulosclerosis, including podocyte foot process effacement, mesangial sclerosis and proteinaceous casts. Podocytes isolated from Tsc2∆podocyte mice show nuclear factor, erythroid derived 2, like 2-mediated increased oxidative stress response on microarray analysis and their autophagic activity is lowered through the mammalian target of rapamycin (mTOR)—unc-51-like kinase 1 pathway. Rapamycin attenuated podocyte dysfunction and extends survival in Tsc2∆podocyte mice. Additionally, mTOR complex 1 (mTORC1) activity is increased in podocytes of renal biopsy specimens obtained from obese patients with chronic kidney disease. Our work shows that mTORC1 hyperactivation in podocytes leads to severe renal dysfunction and that inhibition of mTORC1 activity in podocytes could be a key therapeutic target for obesity-related kidney diseases.

2020-03-21 | GSE106828 | GEO

Drosophila nephrocyte proteome

Project description:Podocyte injury is a major cause of chronic kidney disease, but the involved signaling pathways are largely unknown. Here, we used ultrasensitive proteomics to investigate the response of the native renal podocyte to injury and prioritized candidates by aligning proteinuria in individual mice with protein expression data. The analysis revealed large perturbations of signaling pathways known to associate with FSGS, but also showed perturbation in metabolism and proteins responding to mechanical stress. We employed Drosophila nephrocytes for further functional analysis of selected candidates and studied filtration function and endocytosis. Among the candidates identified was the mechanosensor protein Filamin-B which is a central component of a conserved cellular response to mechanical stress in podocytes in vitro. Loss of Cher, the orthologue of Filamin-B, led to an increased endocytosis in nephrocytes. Of note, Filamin-B expression was only elevated in the Doxorubicin-induced proteome of podocytes but not after LPS induced injury despite the regulation of multiple similar pathways on the systems level. The Filamin-B amount in glomeruli of proteinuric PAN rats correlated with the amount of albumin . In conclusion, this study identified conserved mechanisms of podocyte stress response employing state of the art ultrasensitive proteomics in isolated mouse podocytes and Drosophila nephrocytes.

2020-07-29 | PXD012025 | Pride

Comparison of Glomerular and Podocyte mRNA Profiles in eNOS-/- Diabetic Mice Induced by Streptozotocin

Project description:We compared mRNA profiles of isolated glomeruli versus sorted podocytes between diabetic and control mice. IRG mice crossed with eNOS-/- mice were further bred with podocin-rTTA and TetON-Cre mice to permanently label podocytes before the diabetic injury. Diabetes was induced by injection of streptozotocin. mRNA profiles of isolated glomeruli and sorted podocytes from diabetic and control mice at 10 weeks after induction of diabetes were examined. Consistent with the previous reports, expression of podocyte-specific markers in the glomeruli were down-regulated in the diabetic mice compared to controls. However, these differences disappeared when mRNA levels were corrected for podocyte number per glomerulus. Interestingly, the expression of these markers was not altered in sorted podocytes from diabetic mice, suggesting that the reduced expression of podocyte markers in isolated glomeruli is likely a secondary effect of reduced podocyte number, rather than the loss of differentiation markers. Analysis of the differentially expressed genes in diabetic mice also revealed distinct up-regulated pathways in the glomeruli (mitochondrial function and oxidative stress) and podocytes (actin organization). In conclusion, our data suggest that podocyte-specific gene expression in transcriptome obtained from the whole glomeruli may not represent those of podocytes in the diabetic kidney. We compared mRNA profiles of isolated glomeruli versus sorted podocytes between diabetic and control mice.

2016-03-16 | E-GEOD-79291 | biostudies-arrayexpress

Analysis of podocytes from doxorubicin treated mice

Project description:Podocyte injury is a major cause of chronic kidney disease, but the involved signaling pathways are largely unknown. Here, we used ultrasensitive proteomics to investigate the response of native podocytes from mice with Doxorubicin-induced injury. We observed perturbations of known propagating pathways of glomerular disease such as Jak-Stat signaling and cell adhesion but also in metabolism and proteins responding to mechanical stress. Using a phenotype-guided approach, we prioritized candidates by aligning proteinuria in individual mice with corresponding podocyte protein expression data. The protein Tropomodulin was decreased and correlated negatively with urinary albumin, and Villin and the mechanosensor protein Filamin-B were increased and correlated positively. We employed Drosophila nephrocytes for further functional analysis of prioritized proteins and studied filtration function and endocytosis. Loss of Villin and Tropomodulin led to an impaired function and reduced expression of endocytosis machineries as evidenced by nephrocyte garland proteomes. Loss of Cher, the orthologue of Filamin-B, led to increased endocytosis in nephrocytes, and Filamin-B abundance in single glomeruli correlated with the amount of deposited albumin in the same glomerulus in proteinuric rats. Filamin-B expression could also be increased by applying mechanical stress to podocytes in vitro, but Filamin-B was not increased in podocyte proteomes in LPS-induced injury despite similar regulation of pathways on the global proteome level. In conclusion, this study identified conserved mechanisms of podocyte stress response employing functional proteomics in purified native mouse podocytes and Drosophila nephrocytes.

2020-07-29 | PXD012063 | Pride

Analysis of the podocyte proteome of LPS treated mice

Project description:Podocyte injury is a major cause of chronic kidney disease, but the involved signaling pathways are largely unknown. Here, we used ultrasensitive proteomics to investigate the response of native podocytes from mice with Doxorubicin-induced injury. We observed perturbations of known propagating pathways of glomerular disease such as Jak-Stat signaling and cell adhesion but also in metabolism and proteins responding to mechanical stress. Using a phenotype-guided approach, we prioritized candidates by aligning proteinuria in individual mice with corresponding podocyte protein expression data. The protein Tropomodulin was decreased and correlated negatively with urinary albumin, and Villin and the mechanosensor protein Filamin-B were increased and correlated positively. We employed Drosophila nephrocytes for further functional analysis of prioritized proteins and studied filtration function and endocytosis. Loss of Villin and Tropomodulin led to an impaired function and reduced expression of endocytosis machineries as evidenced by nephrocyte garland proteomes. Loss of Cher, the orthologue of Filamin-B, led to increased endocytosis in nephrocytes, and Filamin-B abundance in single glomeruli correlated with the amount of deposited albumin in the same glomerulus in proteinuric rats. Filamin-B expression could also be increased by applying mechanical stress to podocytes in vitro, but Filamin-B was not increased in podocyte proteomes in LPS-induced injury despite similar regulation of pathways on the global proteome level. In conclusion, this study identified conserved mechanisms of podocyte stress response employing functional proteomics in purified native mouse podocytes and Drosophila nephrocytes.

2020-07-29 | PXD012064 | Pride

OASIS overexpression in murine podocyte cell line

Project description:Podocyte injury is involved in the onset and progression of various kidney diseases. We previously demonstrated that the transcription factor, old astrocyte specifically induced substance (OASIS) in myofibroblasts, contributes to kidney fibrosis, as a novel role of OASIS in the kidneys. Importantly, we found that OASIS is also expressed in podocytes; however, the pathophysiological significance of OASIS in podocytes remains unknown. Upon lipopolysaccharide (LPS) treatment, there is an increase in OASIS in murine podocytes. Enhanced serum creatinine levels and tubular injury, but not albuminuria and podocyte injury, are attenuated upon podocyte-restricted OASIS knockout in LPS-treated mice, as well as diabetic mice. The protective effects of podocyte-specific OASIS deficiency on tubular injury are mediated by protein kinase C iota (PRKCI/PKCι), which is negatively regulated by OASIS in podocytes. Furthermore, podocyte-restricted OASIS transgenic mice show tubular injury and tubulointerstitial fibrosis, with severe albuminuria and podocyte degeneration. Finally, there is an increase in OASIS-positive podocytes in the glomeruli of patients with minimal change nephrotic syndrome and diabetic nephropathy. Taken together, OASIS in podocytes contributes to podocyte and/or tubular injury, in part through decreased PRKCI. The induction of OASIS in podocytes is a critical event for the disturbance of kidney homeostasis.

2022-06-22 | GSE206525 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data