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Establishing a functional response of cardiomyocytes upon prorenin stimulation.

ABSTRACT: Rat neonatal cardiomyocytes have been used to study the effect of (pro-) renin binding upon intracellular messenger systems, hypothesized via IGF2/mannoste 6 phosphate (M6P) -receptors and/or ATP6AP2/(pro)renin receptors, opposed to kidney cells. We have used DNA microarray technology to characterize and compare the expression of over 4800 genes in these cells. Gene expression differences were verified using RT-PCR and indicate the presence of cellular activation upon prorenin bindng, non-overlapping with AT-receptor activation. Exploration and follow-up will lead to further characterization of these pathways. Keywords: loop design

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE4340 | GEO | 2006/08/31

SECONDARY ACCESSION(S): PRJNA94771

REPOSITORIES: GEO

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Project description:Purpose: ATP6AP2 is a single transmembrane protein that has been implicated in regulating various biological process including vascular angiogenesis. However, the exact role of ATP6AP2 during blood vessel development remains largely undefined. Here, we use an inducible endothelial cell (EC)-specific mouse to investigate the role of murine Atp6ap2 gene during both physiological and pathological angiogenesis of the postnatal retina, a well-established model system of angiogenesis. In this study we aimed to Identify what genes are differentially expressed upon loss of PRR(Atp6ap2) in ECs. Methods: ECs were isolated from lungs of both WT and PRR mutant mice at postnatal day 7 (P7). Total RNA was extracted from isolated lung ECs and quantified using Qubit RNA High Sensitivity Assay Kit. RNA integrity was determined using the Bioanalyzer RNA 6000 Nano assay kit. RNA library construction was performed with the TruSeq RNA Library Prep Kit v2 from Illumina. The resulting mRNA library was quantified using Qubit dsDNA High Sensitivity Assay Kit and verified using the Bioanalyzer DNA1000 assay kit. Verified samples were sequenced using the NextSeq 500/550 High Output Kit v2.5 (150 Cycles) on a Nextseq 550 system. Sequenced reads were aligned to the mouse (mm10) reference genome with RNA-seq alignment tool (version 2.0.1). The aligned reads were used to quantify mRNA expression and determine differentially expressed genes using the RNA-seq Differential Expression tool (version 1.0.1). Both alignment and differential expression analysis were performed using the tools in the BaseSpace Sequence Hub. Results: We found 1876 differentially expressed genes of which 823 genes were upregulated while 1053 genes were downregulated. Downregulated genes were enriched in biological processes such as angiogenesis, cell migration and extracellular matrix remodelling, while upregulated genes were enriched in DNA and cell cycle regulation. Conclusions: We identify PRR (ATP6AP2) to play an important role in regulation of the angiogenesis process.

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Establishing a functional response of cardiomyocytes upon prorenin stimulation.

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