Heart gene expression in ANG and salt-induced heart failure
Ontology highlight
ABSTRACT: Balb/cJ and C57BL/6J were subjected to either 0.5µg/kg/min angiotensin II, 3% sodium diet, or the combination. Sham operated animals receiving a 0.3% sodium normal diet were used as controls.
Project description:Balb/cJ and C57BL/6J were subjected to either 0.5µg/kg/min angiotensin II, 3% sodium diet, or the combination. Sham operated animals receiving a 0.3% sodium normal diet were used as controls.
Project description:Balb/CJ mice develop heart failure and quickly decompensate during combination treatment with Angiotensin II and high-sodium diet, while C57BL/6J mice do not. Balb/CJ show a predominantly diastolic heart failure, signs of increased pulmonary pressure, relative sodium and fluid retention, peripheral edema and decompensate with anuria in a cardiorenal syndrome. A F1 cross between the strains have a similar phenotype as C57BL/6J, i.e. is protected. The present study used a second generation backcross of the F1-generation to Balb/CJ and pheotyped a wide variety of renal and circulatory and cardiac traits, identifying 9 new QTL.
Project description:Angiotensin II (Ang II) mediated signaling plays a key role in the development of hypertension associated target organ damages. However, the gene expression changes regulated by Ang II in the early stage of acute cerebral, cardiac, renal, vascular injury remain unclear. we investigated Ang IIâ??mediated gene expression alteration associated with the development of early cerebral, cardiac, renal, vascular injury by microarray assay in a mouse model. All mice were euthanized by an overdose of pentobarbital on days 1, 3 and 7 of Angiotensin II treatment. Total RNA was isolated with TRIzol (Invitrogen) from brains, hearts, kidneys and vessels (n=1-3 per group) at each time point according to manufacturerâ??s instructions. Gene expression profiling was performed using Affymetrix GeneChip mouse Genome 430 2.0 array according to the manufacturerâ??s instructions (Affymetrix, Inc., Santa Clara, CA). On the GeneChip Mouse Genome 430 2.0 Array, over 45,000 probe sets analyze the expression level of over 39,000 transcripts and variants from over 34,000 well characterized mouse genes.
Project description:Transcriptional profiling of suprarenal aorta from ApoE-/- mice (12-14 weeks old, C57BL/6J background) treated by subcutaneous pump with angiotensin II or saline for 7d, 14d and 28d. Includes Ang II-treated samples at 7d found to have dissected aneurysms. Goal was to examine gene expression in developing AAA in this model over time. Experiment Overall Design: Two condition experiment, one suprarenal aorta per array. Saline vs. angiotensin II at 3 time points, with inclusion of 3 Ang II-treated dissected. Total 35 arrays: 6 saline 7d, 6 saline 14d, 5 saline 28d, 4 Ang II 7d, 5 Ang II 14d, 6 Ang II 28d, 3 Ang II-dissected 7d.
Project description:Analysis of angiotensin II effect on left ventricle at gene expression level. The hypothesis tested in the present study was that angiotensin II treatment may affect gene expression in left ventricle in a strain specific manner. Results provide important information about which genes respond to angiotensin II in C57Bl/6N male mice compared to their C57Bl/6J counterparts. Total RNA obtained from isolated left ventricles from C57Bl/6J and C57Bl/6N mice subjected to 48 hours of angiotensin II infusion, via osmotic mini-pumps (500ng/kg/h) implanted sub-cutaneously, compared to sham operated controls.
Project description:We characterized single-cell transcriptional profiles of heart ventricles of C57BL/6J female and male mice subjected to a chronic stress, two weeks of continuous administration of Angiotensin II. The cell preparation we sequenced consisted of metabolically active, nucleated non-myocyte cells which were depleted of endothelial cells, mixed with nuclei isolated from cardiomyocytes. The goal of this experiment included characterizing cellular diversity in stressed and unstressed hearts, uncovering potential drivers of cardiac fibrosis and hypertrophy, and quantifying sexual dimorphism in cardiac gene expression.
Project description:Objective: L-type calcium channels (LTCC) homeostatically regulate calcium on a beat by beat basis, but also provide Ca that over long time scales may contribute to transcriptional regulation. We previously showed that sustained LTCC blockade (CCB) elicits LTCC remodeling in ventricular cardiac myocytes (CM). Here we hypothesize that sustained CCB has broad effects on the expression of genes involved in calcium handling. Methods and Results: Therefore, we subjected adult mice to sustained CCB for 24 hours and performed gene expression profiling. In comparison to vehicle-only control animals, 231 genes were up-regulated, and 111 genes were down-regulated by sustained LTCC blockade (p <0.01). Gene ontology analysis suggested that the CaMKIIdelta signaling pathway was up-regulated in these cells. Unexpectedly, phosphorylation of phospholamban (PLN) at threonine17 (Thr17), an index of CaMKIIdelta activity, was not changed by sustained CCB; however, the degree of phosphorylation of the neighboring PLN-Ser16 substrate site for PKA was significantly reduced by sustained CCB compared to control. Gene expression profiling suggested no change in PKA, but it showed that protein phosphatase 2A (PP2A) mRNA increased, and immunoblots demonstrated that PP2Ac-alpha protein was significantly increased by sustained CCB. Consistent with elevated PP2Ac-alpha protein expression LTCC exhibited decreased phosphorylation of the C-terminal Ser1928 PKA substrate site. Conclusions: We conclude that sustained CCB elicits a spectrum of transcriptional events, including compensatory up-regulation of LTCC and PP2Ac-alpha. Although this study is restricted to mouse, these results suggest the new hypothesis that clinically-relevant sustained LTCC blockade in humans results in changes in gene regulation in the heart. Keywords: L-type calcium channel, calcium channel blockade, verapamil Female and male ICR mice (12-14 weeks age) weighing between 25 and 30 grams were anesthetized with a ketamine/xylazine mixture ( i.p.) allowing the subcutaneous implantation of miniosmotic pumps (Alzet, model 2001). The pumps were filled with either verapamil or vehicle (0.02% ascorbic acid). Control animals carried mini-pumps with vehicle and control animals were investigated in parallel with each set of experimental animals. Mini pumps delivered verapamil at 3.6 mg/kg/day for 24 (RNA)-48 (protein) hours. After treatment animals were anesthetized and weighed. Hearts were excised, rinsed, blotted dry, weighed, and then frozen on dry ice and the stored at -80oC until studied. Animals were anesthetized and euthanized according to animal protocols approved by the University of Kentucky Institutional Animal Care and Use Committee. This investigation conforms with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication NO. 85-23, revised 1996). Left ventricular free wall from female mice was rapidly excised and either snap frozen at -80oC or used immediately for RNA isolation. Three VER treated mice and 3 vehicle treated mice were used to generate RNA for microarray. Total RNA was isolated using the RNAqueous -4PCR kit (Ambion) and quantitated spectrophotometrically at 260nm. Contaminating genomic DNA was eliminated by DNase treatment (Ambion). RNA quality was assessed using the Agilent 2100 Bioanalyzer. Microarray data was obtained using the Affymetrix 430 V2 GeneChip (representing 45,101 probe sets), in accordance with the manufacturer’s specifications.
Project description:This experiment compares gene expression in kidneys of control and ACTH treated mice. <br>Kidney RNA was prepared from 6 mice: three controls and three ACTH treated and processed by the microarray team at Ark Genomics and the Roslin Institute. <br>RNAs were processed through standard Affymetrix protocols, with one round of cDNA amplification. Processed RNAs were hybridised to Affymetrix Mouse Genome 430 2.0 GeneChip, and data were extracted through the GCOS software. <br>CEL files were made available for further data processing via RMA in Bioconductor.
Project description:Angiotensin II stimulates fibroblast proliferation and substantially alters gene expression patterns leading to cardiac remodeling, but the mechanisms for such differences are unknown. MicroRNAs are a novel mechanism for gene expression regulation. Herein we tested the miRNAs patterns in mouse heart using microarray assay, and investigated their role in angiotensin II-induced cardiac remodeling. In this study, nine samples (three groups) from mouse were used to acquire the miRNA expression profiling and the function of the abudant miRNAs in the heart were analyzed by bioinformatic methods.