Expression data from mDCT cell-line over-expressing hMR
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ABSTRACT: Target gene of mineralocorticoid receptor (MR) is comparatively unknown, although distal convoluted tubule (DCT) expresses MR in in vivo. We used microarray and immortalized murine DCT cell-line overexpressing human MR with treatment of aldosterone to elucidate target genes of MR in DCT. mDCT overexpresses human MR by lipofection and is treated for 3 hours with ethanol (g1), or 10^-9 M aldosterone (g2), or 10^-7 M aldosterone (g3), or 10^-7 M aldosterone with pretreatment of 5 x 10^-6 M spironolactone (g4) for 2 hours.
Project description:Target gene of mineralocorticoid receptor (MR) is comparatively unknown, although distal convoluted tubule (DCT) expresses MR in in vivo. We used microarray and immortalized murine DCT cell-line overexpressing human MR with treatment of aldosterone to elucidate target genes of MR in DCT.
Project description:Kidney distal convoluted tubules (DCT) are important for regulation of urinary salt excretion. Aldosterone is known to exert long-term effects in this segment, and regulate sodium reabsorption via increasing abundance of the Na+-Cl- cotransporter (NCC) and the epithelial Na channel ENaC. Whether acute effects of aldosterone occur in the DCT and the potential signaling networks remain unknown. Here in this study, we aim to identify the acute aldosterone-mediated signaling (rapid effects) in the DCT.
Project description:The “aldosterone paradox” is a poorly understood physiological mechanism in which increases in the hormone aldosterone either serve to stabilize blood pressure by increasing sodium-chloride (NaCl) reabsorption or to excrete potassium (K+). Under low NaCl intake, abundance and activity of the thiazide-sensitive NaCl cotransporter NCC, expressed in the distal convoluted tubule (DCT), is high, whereas it is low during high K+ intake - leading to increased delivery of NaCl to downstream segments and increased electrogenic K+ secretion. Despite a critical role in blood pressure and K+ homeostasis, little is known about the molecular alterations in the DCT during the aldosterone paradox, and how they control NCC levels. The objective of this study was to define the proteome of the DCT and how it is modified by increased circulating aldosterone levels subsequent to long-term low dietary NaCl or high K+ intake.
Project description:Aldosterone, the main mineralocorticoid hormone in Humans, has a major role in maintaining the hemo-electrolytique homeostasis, by acting in the distal part of nephron. This steroid hormone mainly acts through its binding to a ligand-induced transcription factor, the mineralocorticoid receptor (MR). MR binds to specific genomic sequences, where it recruits transcriptional coregulators, to activate or repress target gene transcription. The aim of this work was to access the whole aldosterone-dependand MR target genes, by comparing sequenced data from aldosterone or vehicle-treated samples. Anti-MR ChIP-seq in Human Kidney GFP-hMR cells treated with vehicle or Aldosterone.
Project description:The steroid hormone aldosterone plays a role in vascular function and disease. Aldosterone activates the mineralocorticoid receptor (MR), a ligand-activated transcription factor. MR have been found to be expressed in vascular cells and vessels. We used microarrays to identify the global programme of gene expression changes in mouse aortas treated with vehicle (DMSO) or aldosterone. Wild type C57Bl6 male mice were treated with spironolactone (20 mg/kg/day, an MR antagonist) for 5 days by Sub-cutaneous pellet to suppress basal MR-mediated gene expression. Aortas were then harvested and treated ex vivo in organ culture with vehicle or 100 nM aldosterone for 2, 4, or 8 hours. Three aortas were pooled for each timepoint and treatment, and three pooled biological replicates were performed.
Project description:The late-gestation fetal lung has relatively high levels of expression of the mineralocorticoid receptor (MR) as well as the glucocorticoid receptor (GR), suggesting that endogenous corticosteroids may act in the lung through binding at MR as well as GR. This study was designed to determine the effects of physiologically relevant increases in steroids on MR and GR in the late-gestation lung. The GR agonist, betamethasone, the MR agonist, aldosterone, or both agonists were infused intravenously for 48 hours in ovine fetuses of approximately 130 days gestation. Effects on airway pressures during stepwise inflation of the in situ lung, expression of ENaC and Na,K ATPase, and elastin and collagen content were determined at the end of the infusions. We found that aldosterone significantly reduced the initial airway pressures measured in situ during inflation. This effect did not occur with betamethasone alone or in combination with aldosterone. Conversely, betamethasone, but not aldosterone, significantly increased expression of the epithelial sodium channel (ENaC) subunit mRNAs, and collagen and elastin content in the lungs. Aldosterone altered novel gene pathways in the fetal lung, suggesting effects on lung compliance via genes which influence immune pathways and lung stiffness. The results are consistent with corticosteroid-induced fluid reabsorption at birth through GR rather than MR, but suggest that MR contributes effects facilitating lung inflation with the first breaths via nonclassical mechanisms. 4 sets of twin sheep fetuses at approximately 130d gestation (term is 147d) were used. One twin was infused with 0.2 mg aldosterone for 48 h, the other received vehicle.
Project description:The mineralocorticoid receptor is expressed in the rat and human retina. We previously showed that intravitreal injection of aldosterone in rat eyes induced retinal œdème and choroidal vasodilation and permeability through regulation of ion/water channels (Zhao et al. Faseb J, 2009; Zhao et al. J Clin Invest 2012). Illicit activation of MR induces inflammation, oxidative stress and tissue remodeling in cardiovascular and renal diseases independent of hypertension. We performed a full transcriptomic study destinated to identify genes regulated by aldosterone in the whole retina of rat.
Project description:To test for a function effect of mineralocorticoid receptor modulation in skeletal muscle, global gene expression analysis was conducted on human myltubes treated with a mineralocorticoid receptor agonist or antagonist. 9 total samples were analyzed. 3 biological replicates for each of the following treatment groups were included: 1) aldosterone; 2) spironolactone; 3) vehicle (ethanol). Pairwise comparison between groups were carried out and a fold-change ≥2 were selected.
Project description:To elucidate the aldosterone-regulated genes in vascular endothelial cells, we developed human endothelial cell line (EAhy926) stably expressing human mineralocorticoid receptor by retroviral system (MR-EAhy). Then gene expression profile of MR-EAhy stimulated with or without aldosterone was compared using DNA microarray analysis. MR-EAhy were serum-deprived for 24h with 0.2% dextran-coated charcoal-treated FBS, and then stimulated with or without aldosterone (10-9 M) for 6 hr. Then cells were harvested for RNA extraction.
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