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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiles of the adult mesangial cell (GUDMAP Series_id: 37)

ABSTRACT: The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing kidney. The central thesis is straightforward. The combination of fluorescent activated cell sorting (FACS) plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing kidney. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene in FACS isolated components of the developing kidney. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. Meis1-GFP BAC transgenic mice were utilized to isolate the mesangial cells from adult kidneys. The mesangial cells were isolated from the glomerulus using collagenase digestion, differential sieving, trypsin treatment and FACS. The RNA was isolated from purified mesangial cells and the gene expression profiles were determined by microarrays.

ORGANISM(S): Mus musculus

SUBMITTER: GUDMAP Developers

PROVIDER: E-GEOD-20687 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RNA-SEQ analysis of various developing and adult kidney components.

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RNA-SEQ analysis of the developing lower urogenital region

Project description:We will use micro-dissection and FACS techniques to isolate cell types from the developing lower urogenital region and perform a transcriptional analysis using RNA-SEQ. This analysis will determine the transcriptional profile of each cell type, identify compartment specific transcripts, compartment specific transcript isoforms and cell-type specific long-noncoding RNAs. In addition the unbiased nature of RNA-SEQ will potentially identify novel transcripts that have not been annotated in the database. Total RNA was obtained from FACS isolated embryonic day 17.5 bladders using Tg(Upk2-cre)6Xrw x Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo transgenic mice. The long term goal is to generate a transcriptional atlas of developing lower urogenital region.

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Gene expression profiles of the E15.5 developing podocyte (GUDMAP Series_id: 33)

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2010-07-16 | E-GEOD-17145 | biostudies-arrayexpress

Gene expression profiles of components of the developing kidney (GUDMAP Series_id: 22)

Project description:The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing kidney. The central thesis is straightforward. The combination of fluorescent activated cell sorting (FACS) plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing kidney. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene in FACS isolated components of the developing kidney. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. Keywords: Comparison of kidney components. At different developmental time points we isolate discrete elements of the kidney by using fluorescent activated cell sorting (FACS) and then define their gene expression profiles with microarrays.

2008-08-04 | E-GEOD-12309 | biostudies-arrayexpress

Comparative phosphoproteomic analysis of mammalian renal glomeruli

Project description:Glomerular biology is dependent on tightly controlled signal transduction networks that control phosphorylation of signaling proteins such as cytoskeletal regulators or slit diaphragm proteins of kidney podocytes. Recent, in-depth analysis of the mouse glomerular phosphoproteome by tandem mass spectrometry confidently identified thousands of phosphorylation sites providing a unique opportunity to study signaling in vivo. However, the multitude of phosphorylated residues demands for additional systems biology approaches to separate physiologically important sites from non-functional phosphorylated residues. Cross-species comparison of phosphorylation events is a powerful mean to functionally prioritize and identify physiologically meaningful phosphorylation sites. Here, we present the result of additional phosphoproteomic analyses of the glomerular phosphoproteome in cow and rat to allow cross-species comparisons. We discovered several phosphorylation sites with potentially high biological relevance, e.g. tyrosine phosphorylation of the cytoskeletal regulator synaptopodin and the slit diaphragm protein neph-1 (Kirrel). Moreover, cross-species comparisons revealed conserved phosphorylation of the slit diaphragm protein nephrin on an acidic cluster at the intracellular terminus and conserved podocin phosphorylation on the very carboxyl terminus of the protein. Given the pivotal role of podocin for glomerular biology we studied a highly conserved podocin phosphorylation site in greater detail and show that phosphorylation regulates affinity of the interaction with nephrin and CD2AP. Taken together, these results suggest that species comparisons of phosphoproteomic data may reveal regulatory principles in glomerular biology.

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