Dataset Information

Drosophila nephrocyte proteome

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Pericardial Nephrocyte

DISEASE(S): Focal Segmental Glomerulosclerosis

SUBMITTER: Markus Rinschen

LAB HEAD: Markus Rinschen

PROVIDER: PXD012025 | Pride | 2020-07-29

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20180329_Q2_AWH_ColID_562_ProjID_1108_Benzing_sample_1.raw	Raw
	20180329_Q2_AWH_ColID_562_ProjID_1108_Benzing_sample_10.raw	Raw
	20180329_Q2_AWH_ColID_562_ProjID_1108_Benzing_sample_11.raw	Raw
	20180329_Q2_AWH_ColID_562_ProjID_1108_Benzing_sample_12.raw	Raw
	20180329_Q2_AWH_ColID_562_ProjID_1108_Benzing_sample_13.raw	Raw

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Publications

Proteome Analysis of Isolated Podocytes Reveals Stress Responses in Glomerular Sclerosis.

Koehler Sybille S Kuczkowski Alexander A Kuehne Lucas L Jüngst Christian C Hoehne Martin M Grahammer Florian F Eddy Sean S Kretzler Matthias M Beck Bodo B BB Höhfeld Jörg J Schermer Bernhard B Benzing Thomas T Brinkkoetter Paul T PT Rinschen Markus M MM

Journal of the American Society of Nephrology : JASN 20200211 3

<h4>Background</h4>Understanding podocyte-specific responses to injury at a systems level is difficult because injury leads to podocyte loss or an increase of extracellular matrix, altering glomerular cellular composition. Finding a window into early podocyte injury might help identify molecular pathways involved in the podocyte stress response.<h4>Methods</h4>We developed an approach to apply proteome analysis to very small samples of purified podocyte fractions. To examine podocytes in early d ...[more]

PMID: 32047005

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