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- W428319672 abstract "Proteomic studies of late distal convoluted tubule and connecting tubule have so far been hindered by insufficient yield and purity of manually isolated tubular cells. Our aim was to determine the effects of 24-hours elevated plasma aldosterone on the protein expression profile in these tubules. Mice endogenously expressing enhanced green fluorescent protein in the late distal convoluted tubule, connecting tubule, and initial cortical collecting ducts were treated with a single subcutaneous injection of 2.0 mg/kg aldosterone or vehicle. Before the sacrifice, the plasma aldosterone was 5.6-fold increased in the aldosterone treated mice. Suspensions of single viable kidney cells were subjected to fluorescence activated cells sorting, yielded 1.4 million cells per mouse with over 69% purity. Three biological replicates of pooled protein samples from 10 mice (100 μg) in each vehicle or aldosterone treated group were digested with trypsin and labelled with 8-plex iTRAQ reagents. Strong cation exchange yielded 18 fractions for liquid chromatography tandem mass spectrometry. The resulting spectra were matched to peptides by the SEQUEST search algorithm, which identified 3904 peptides corresponding to 506 unique proteins. A change in abundance was observed in several of these proteins after 24-hour aldosterone treatment. Among the increased proteins, pyruvate dehydrogenase E1 and pyruvate kinase isozyme M2 were confirmed by immunohistochemistry. These enzymes are involved in the carbohydrate metabolism, suggesting an increased capacity for energy production after 24-hours aldosterone treatment." @default.
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- W428319672 date "2012-04-01" @default.
- W428319672 modified "2023-10-12" @default.
- W428319672 title "Identification of proteins regulated by 24‐hour aldosterone treatment in late distal convoluted tubules, connecting tubules and initial cortical collecting ducts" @default.
- W428319672 doi "https://doi.org/10.1096/fasebj.26.1_supplement.885.9" @default.
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