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Gene expression analysis of rat glomerular parietal epithelial cells

ABSTRACT: Very little is known about the function of glomerular parietal epithelial cells (PECs). In this study, we performed genome-wide expression analysis on PEC-enriched capsulated vs. PEC-deprived decapsulated rat glomeruli to determine the transcriptional state of PECs under normal conditions. We identified hundreds of differentially expressed genes that mapped to distinct biologic modules including development, tight junction, ion transport, and metabolic processes. Since developmental programs were highly enriched in PECs, we characterized several of their candidate members at the protein level. Collectively, our findings confirm that PECs are multifaceted cells and help define their diverse functional repertoire.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE54085 | GEO | 2014/08/01

SECONDARY ACCESSION(S): PRJNA235345

REPOSITORIES: GEO

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Gene expression analysis of rat glomerular parietal epithelial cells

Project description:Very little is known about the function of glomerular parietal epithelial cells (PECs). In this study, we performed genome-wide expression analysis on PEC-enriched capsulated vs. PEC-deprived decapsulated rat glomeruli to determine the transcriptional state of PECs under normal conditions. We identified hundreds of differentially expressed genes that mapped to distinct biologic modules including development, tight junction, ion transport, and metabolic processes. Since developmental programs were highly enriched in PECs, we characterized several of their candidate members at the protein level. Collectively, our findings confirm that PECs are multifaceted cells and help define their diverse functional repertoire. We developed a sequential sieving methodology to selectively isolate capsulated (PEC-enriched) glomeruli from un-capsulated (PEC-deprived) glomeruli in rats. Three samples were obtained for each preparation. Total RNA was isolated and its integrity confirmed via Bioanalyzer 2100. Each sample (n =6) was hybridized to an Affymetrix Rat GeneChip 1.0ST microarray.

2014-08-01 | E-GEOD-54085 | biostudies-arrayexpress

Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin

Project description: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. Total RNA for genechip analysis was derived from: 1) Primary parietal cells isolated by FACS sorting of glomeruli cultivated in 1a) RPMI media 1b) EGM-MV media 2) Primary podocyte cells isolated by FACS sorting of glomeruli cultivated in 2a) RPMI media 2b) EGM-MV media

2011-11-16 | E-GEOD-33714 | biostudies-arrayexpress

Colony stimulating factor-1 receptor drives glomerular parietal epithelial cell activation in focal segmental glomerulosclerosis

Project description:Parietal epithelial cells (PECs) are part of renal progenitor cells with similarities to bone marrow stem cell niche. In focal segmental glomerulosclerosis (FSGS) PECs become activated and contribute to extracellular matrix deposition. Colony stimulating factor-1 (CSF-1), a hematopoietic growth factor, acts via its specific receptor, CSF-1R, and has been implicated in several glomerular diseases, although its role on PEC activation is unknown. We found that CSF-1R was upregulated in PECs and podocytes from human biopsies with FSGS. Through in vitro studies, we demonstrated that PECs constitutively express CSF-1R. Incubation with CSF-1 induced CSF-1R upregulation and significant transcriptional regulation of genes involved in pathways associated with PEC activation. Specifically, CSF-1/CSF-1R activated the ERK1/2 pathway and upregulated CD44 in PECs, while both ERK and CSF-1R inhibitors reduced CD44 expression. Our functional studies showed that CSF-1 induced PEC proliferation and migration, while reducing the differentiation of PECs into podocytes. These results were validated in the Adriamycin-induced FSGS experimental model. Importantly, treatment with either the CSF-1R-specific inhibitor GW2580 or Ki20227 provided a robust therapeutic effect. In conclusion, we provide the first evidence of the role of the CSF-1/CSF-1R pathway in PEC activation in FSGS, paving the way for future clinical studies investigating the therapeutic effect of CSF-1R inhibitors on FSGS in humans.

2024-02-13 | GSE255489 | GEO

Primary cultures of glomerular parietal epithelial cells or podocytes with proven origin

2011-11-16 | GSE33714 | GEO

Identification of the cell subset generating crescents within human parietal epithelial cells (PECs)

Project description:We generated a single-cell RNA sequencing (scRNAseq) dataset of primary cultures of human glomerular parietal epithelial cells. Comparing this dataset with an already available dataset obtained by scRNAseq of human renal progenitor cells (GSE137620), we identified an immature PEC subset characterized by the expression of stem cell markers and stratifin (SFN). Moreover, we analyzed the effect of treatment with the HDAC inhibitor panobinostat on PEC transcriptome demonstrating the upregulation of podocyte-specific markers in different PEC subsets including the stratifin-expressing subset.

2022-09-24 | GSE195797 | GEO

WT1+ parietal epithelial progenitor cells are essential for renal proximal tubule repair and regeneration

Project description:Purpose: We have found that WT1+ parietal epithelial progenitor cells contribute to renal proximal tubule repair and regeneration by cell lineage tracing and direct differentiation analysis, but the transcription profile of these WT1+ PECs is largely unkown. Here, we aimed to unveil the transcriptional features of WT1+ PECs through single-cell RNA sequencing (scRNA-seq). Methods: Single cell suspension was prepared from kidney cortex of WT1CreERT2; Rosa26-tdTfl/+ mice that underwent sham or ischemic reperfusion injury (IRI) 24h .TdT+ cells were enriched by fluorescence activated cell sorter (FACS) and further analyzed with BD Rhapsody platform. Results: After quality control and cell filtering, 26,759 cells were informative (Dataset S1) and divided into 10 clusters using Harmony algorithm to reduce batch effects. GO analysis and QuSAGE heatmap suggested that WT1+ PECs might promote kidney regeneration after AKI through TGF-β and Notch signaling pathway. Marker gene expression analysis showed that WT1+ PECs shared the characteristics of STCs. Pseudotemporal analysis indicated that that WT1+ PECs developed to PTECs through STCs intermediate stage. Conclusions: scRNA-seq indicated that the rare, quiescent PECs supplied PTECs through STCs intermediate stage after severe ischemic injury, and this process most likely was regulated through TGF-β and Notch signaling pathway.

2023-03-22 | GSE184601 | GEO

Expression data of metformin treated wild type and parietal cell- specific AMPK-deficient mice

Project description:Genes related to AMPK activation, cellular respiration, and metabolism are enriched in the gastric parietal cell population. Metformin is known activator of AMPK. We used microarray analysis to identify metformin targets in mature parietal cells and in parietal cell progenitors.

2020-04-01 | GSE141673 | GEO

Visceral Vs. Parietal Mesothelia

Project description:The gene expression profiles of murine visceral and parietal mesothelia were analyzed by microarray. Visceral and parietal mesothelia were isolated from the same adult Wt1-cre;Rosa26ReYFP reporter mice and analyzed. Whole tracscriptome expression analysis of total RNA was performed using Affymetrix mouse Gene 1.0 ST arrays.

2012-05-12 | E-MTAB-1399 | biostudies-arrayexpress

Single-cell RNA sequencing reveals the mesangial identity and species diversity of glomerular cell transcriptomes

Project description:Molecular characterization of the individual cell types in human kidney as well as model organisms are critical in defining organ function and understanding translational aspects of biomedical research. Previous studies have uncovered gene expression profiles of several kidney glomerular cell types, however, important cells, including mesangial (MCs) and glomerular parietal epithelial cells (PECs), were missing or incompletely described, and a systematic comparison between mouse and human kidney is lacking. To this end, we used Smart-seq2 to profile 4332 individual glomerulus-associated cells isolated from human living donor renal biopsies and mouse kidney. The analysis revealed genetic programs for all four glomerular cell types (podocytes, glomerular endothelial cells, MCs and PECs) as well as rare glomerulus-associated macula densa cells (MDCs). Importantly, we detected heterogeneity in glomerulus-associated Pdgfrb-expressing cells, including bona fide intraglomerular MCs with the functionally active phagocytic molecular machinery, as well as a unique mural cell type located in the central stalk region of the glomerulus tuft. Furthermore, we observed remarkable species differences in the individual gene expression profiles of defined glomerular cell types that highlight translational challenges in the field and provide a guide to design translational studies.

2020-11-21 | GSE160048 | GEO

Col4a3 mouse glomerular datasets

Project description:Glomeruli were isolated from Col4a3-/- and Ctrl mice. The extracellular matrix was enriched and both cellular and matrix fraction were analysed by mass spectrometry.

2022-08-12 | PXD022392 | Pride

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