Dataset Information

Comparative protein profiling of wild type and zyxin knockout podocytes

ABSTRACT: Podocyte detachment due to mechanical stress is a common issue in hypertension-induced kidney disease. The present study investigated the role of zyxin in mechanically stretched podocytes, a model for glomerular hypertension. Conditionally immortalized differentiated podocytes were seeded on flexible silicon membranes of a six well plate, which was mounted on a manifold connected to a custom-built stretch apparatus (NIPOKA GmbH, Greifswald, Germany), which induced cyclic pressure variations resulting in upward and downward motion of the silicone membranes. We found that zyxin is essential for the adhesion of the postmitotic cell type, the podocytes, in vitro during mechanical stretch as well as in vivo for the maintenance of the kidney filtration.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Glomerular Visceral Epithelial Cell

SUBMITTER: Elke Hammer

LAB HEAD: Elke Hammer

PROVIDER: PXD050861 | Pride | 2024-04-30

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20211209_Exp_AW_Anat_Ctrl_KO_1.raw	Raw
	20211209_Exp_AW_Anat_Ctrl_KO_12.raw	Raw
	20211209_Exp_AW_Anat_Ctrl_KO_24.raw	Raw
	20211209_Exp_AW_Anat_Zyx_KO_24_17.raw	Raw
	20211209_Exp_AW_Anat_Zyx_KO_24_2.raw	Raw

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Publications

Zyxin is important for the stability and function of podocytes, especially during mechanical stretch.

Kliewe Felix F Siegerist Florian F Hammer Elke E Al-Hasani Jaafar J Amling Theodor Rolf Jakob TRJ Hollemann Jonas Zeno Eddy JZE Schindler Maximilian M Drenic Vedran V Simm Stefan S Amann Kerstin K Daniel Christoph C Lindenmeyer Maja M Hecker Markus M Völker Uwe U Endlich Nicole N

Communications biology 20240411 1

Podocyte detachment due to mechanical stress is a common issue in hypertension-induced kidney disease. This study highlights the role of zyxin for podocyte stability and function. We have found that zyxin is significantly up-regulated in podocytes after mechanical stretch and relocalizes from focal adhesions to actin filaments. In zyxin knockout podocytes, we found that the loss of zyxin reduced the expression of vinculin and VASP as well as the expression of matrix proteins, such as fibronectin ...[more]

PMID: 38605154

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Project description:Acute respiratory distress syndrome (ARDS) is a catastrophic form of acute lung injury (ALI). The necessity for mechanical ventilation (MV) renders patients at risk for ventilator induced lung injury (VILI). Exposure to repetitive cyclic stretch (CS) and/or over-inflation exacerbates injury. Reducing tidal volume (VT) is the only therapeutic strategy shown to mitigate morbidity and mortality. Cyclic stretch has been shown to differentially regulate gene expression in part through the activation of mammalian mitogen-activated protein kinase (MAPK). Although these studies have shown both molecular and cellular alterations, no unifying hypothesis to explain MV-induced lung injury has emerged. In the current study, we hypothesized that coordinated expression of cyclic stretch (CS)-responsive genes relies on the presence of common CS-sensitive regulatory elements. To identify CS-responsive genes, we undertook a comparative examination of the gene expression profile of human bronchial epithelial airway (Beas-2B) cells in response to various injurious stimuli involved in the pathogenesis of acute lung injury (ALI)/Ventilator induced lung injury (VILI): cyclic stretch, tumor necrosis factor alpha (TNF-a), and lipopolysaccharide (LPS). Experiment Overall Design: Human Bronchial Epithelial Cells (Beas-B2) cells grown on silicon elastic plates coated with Type I collagen (Flexercell International, McKeesport, PA) were exposed to six regiments for 4 h: 1) control (static, [control]); 2) mechanical stretch (25 PKa, 30 cycles per min, [stretch]); 3) LPS (1 mcg/ml [LPS]); 4) TNF-Î± (20 ng/ml; [TNF]); 5) mechanical stretch plus LPS [LPS+S], and 6) mechanical stretch plus TNF-Î± [TNF+S]. Total RNA (duplicate experiments) was extracted using TRIZOL reagent (as per manufactures specifications) and purified using Qiagen mRNA purification Kit (as per manufacturers specifications). mRNA was hybridized to Affymetrix Human U133plus2.0 chips. Probe based analysis, background reduction, and quantile data normalization was performed in MeV 4.0 of TM4 using Robust Multi-array Average (RMA).