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Enriched Single-Nucleus RNA-Sequencing Reveals Unique Attributes of Distal Convoluted Tubule Cells

ABSTRACT: Enriched Single-Nucleus RNA-Sequencing Reveals Unique Attributes of Distal Convoluted Tubule Cells

PROVIDER: PRJNA949574 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23989615_1.fastq.gz	Fastqsanger.gz
	SRR23989615_2.fastq.gz	Fastqsanger.gz
	SRR23989616_1.fastq.gz	Fastqsanger.gz
	SRR23989616_2.fastq.gz	Fastqsanger.gz
	SRR23989617_1.fastq.gz	Fastqsanger.gz

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Enriched Single-Nucleus RNA-Sequencing Reveals Unique Attributes of Distal Convoluted Tubule Cells

Project description:Distal convoluted tubule (DCT) cells display a remarkable ability to adapt to the dietary potassium intake. We applied single-nucleus RNA-seq to map the plasticity of enriched mouse DCT cells in response to dietary potassium restriction

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SUMOylation proteomic analysis of renal distal convoluted tubule cells

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Identification of renal distal convoluted tubules enriched genes

Project description:The renal distal convoluted tubule DCT is a short part of the distal nephron with specific functions transporting ions and thereby modifying Na, Ca, Mg, K balance A transcriptomic analysis of the DCT was performed to identify specific gene expression which would implicate those genes in specific DCT function Fluorescent DCT (EGFP+) were sorted out from a renal tubule suspension by a COPAS (Complex Object Parametric analyser and sorter). EGFP- and the total (ALL) fractions were also sorted by COPAS. Gene expression for each fractions (EGFP+, EGFP-, ALL) was performed in 4 mice

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Identification of renal distal convoluted tubules enriched genes

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Project description:The kidney distal convoluted tubule (DCT) plays an important role in body sodium regulation and thus control of blood pressure. The main sodium reabsorption pathways in the DCT are the epithelial sodium channel (ENaC) and the thiazide-sensitive NaCl cotransporter (NCC), the functions of which can be modulated by the hormone vasopressin (VP) acting via uncharacterized signaling cascades. We performed large scale stable isotope labeling by amino acids in cell culture (SILAC) based quantitative phosphoproteomics of cultured mouse DCT cells (mpkDCT) to map global changes in protein phosphorylation events upon acute treatment with the VP type II receptor agonist 1-desamino-8-D-arginine vasopressin (dDAVP). The aim of this study is to identify unique VP signaling cascades in DCT cells that may be important for regulating ENaC or NCC activity. These pathways may be novel targets for modulation of blood pressure.

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Comparative proteomic analysis of kidney distal convoluted tubule and cortical collecting duct cells following long-term hormonal stimulation

Project description:The distal convoluted tubule (DCT) and the cortical collecting ducts (CCD) are portions of renal tubule that are partly responsible for maintaining the systemic concentrations of potassium, sodium, calcium and magnesium. Despite being structurally similar, DCT and CCD cells have different transport capabilities due to a variety of different membrane-associated transport proteins. However, DCT and CCD cells appear to be modulated via the same hormones. The objective of this study was assess the differential response of DCT and CCD cells to long-term exposure to the hormones vasopressin or angiotensin II, both of which modulate DCT and CCD cells differently. Mass spectrometry based quantitative proteomics was used to profile the differential proteome between DCT and CCD.

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JeB001: Elucidation of the distal convoluted tubule transcriptome identified new candidate players in magnesium balance

Project description:The distal convoluted tubule (DCT) plays a key role in the fine-tuning of renal Mg2+ reabsorption. In recent years, hereditary magnesium (Mg2+) transport disorders have helped to identify important players in DCT Mg2+ homeostasis. Here, we report the Mg2+-sensitive transcriptional expression of the DCT transcriptome using gene expression microarray analysis. Mice expressing eGFP under a DCT-specific promoter were subjected to Mg2+-deficient or Mg2+-enriched diets. Subsequently, the Complex Object Parametric Analyzer and Sorter (COPAS) allowed isolation of eGFP-positive DCT cells. RNA extracts were subjected to pair-wise microarray analysis, high vs. low Mg2+. Of all genes in the genome, 46 were shown to be differentially expressed with a minimal fold-change of 2. Several known magnesiotropic genes, such as Trpm6 and Parvalbumin, were upregulated in the low Mg2+ fraction. Moreover, new genes were identified that are potentially involved in renal Mg2+ homeostasis. To confirm that the selected candidate genes are regulated by dietary Mg2+ availability, the expression levels of Slc41a3, Pcbd1, Tbc1d4 and Umod were determined by RT-PCR analysis. Indeed, all four genes were shown to be significantly upregulated in mice fed the Mg2+-deficient diet. In conclusion, by elucidating the Mg2+-sensitive DCT transcriptome, new candidate players in renal Mg2+ homeostasis were identified. RNA samples were isolated from PV-eGFP-positive tubules from mice fed on a low (L-samples) or high (H-samples) magnesium diet. Littermate H and L samples were labeled and put on mouse microarrays together. Two of the four sample sets were analyzed in dye-swap to compensate for dye-biases.

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Acute effects of Salbutamol in Mouse Kidney Distal Convoluted Tubules (mpkDCT) Cells

Project description:The sympathetic nervous system (SNS) plays a central role in blood pressure regulation. Recent studies have shown that cells of the distal convoluted tubule (DCT) can be stimulated directly via the beta-adrenergic receptor resulting in activation of the NaCl cotransporter NCC. Whether these effects are mediated by the beta1- or beta2-adrenergic receptor is unclear, and the acute signaling cascades rapidly activated by beta-adrenergic receptor stimulation in the DCT are unknown. Here in this study, we aim to identify the rapid salbutamol-mediated (beta2-adrenergic receptor) signaling in the DCT by looking at global protein phosphorylation changes in the mpkDCT cells.

2021-05-17 | PXD019051 | Pride

JeB001: Elucidation of the distal convoluted tubule transcriptome identified new candidate players in magnesium balance

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