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Transcriptome profiling of wildtype, nonoperated mouse hearts compared to wildtype, sham operated hearts

ABSTRACT: The purpose of the present study was to investigate the activation of the cardiac inflammatory response in the hearts of SHAM (surgical opening of the chest) mice in comparison to CONTROL hearts isolated from naïve (not operated) mice. Male C57BL/6J (10 weeks) were subjected to an open-chest ischemia-reperfusion (I/R) SHAM procedure in order to examine the effects of this surgery alone in producing cardiac inflammation. The procedure consisted of thoracotomy and a suture passed around the left anterior descending artery (LAD). The mouse chest was kept open for approximately 45 min. We chose to study an open-chest I/R SHAM because this is the reference surgery for the most common surgical procedure used to study cardiac inflammation. Mice were sacrificed 5 days after the procedure and the hearts were removed and used in RNAseq experiments.

ORGANISM(S): Mus musculus

PROVIDER: GSE120867 | GEO | 2020/10/04

REPOSITORIES: GEO

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Project description:Accurate and comprehensive de novo transcriptome profiling in heart is a central issue to better understand cardiac physiology and diseases. Although significant progress has been made in genome-wide profiling for quantitative changes in cardiac gene expression, current knowledge offers limited insights to the total complexity in cardiac transcriptome at individual exon level. To develop more robust bioinformatic approaches to analyze high-throughput RNA sequencing (RNA-Seq) data, with the focus on the investigation of transcriptome complexity at individual exon and transcript levels. In addition to overall gene expression analysis, the methods developed in this study were used to analyze RNA-Seq data with respect to individual transcript isoforms, novel spliced exons, novel alternative terminal exons, novel transcript clusters (i.e., novel genes) and long non-coding RNA genes. We applied these approaches to RNA-Seq data obtained from mouse hearts following pressure-overload induced by trans-aortic constriction. Based on experimental validations, analyses of the features of the identified exons/transcripts, and expression analyses including previously published RNA-Seq data, we demonstrate that the methods are highly effective in detecting and quantifying individual exons and transcripts. Novel insights inferred from the examined aspects of the cardiac transcriptome open ways to further experimental investigations. Our work provided a comprehensive set of methods to analyze mouse cardiac transcriptome complexity at individual exon and transcript levels. Applications of the methods may infer important new insights to gene regulation in normal and disease hearts in terms of exon utilization and potential involvement of novel components of cardiac transcriptome. Left ventricular tissues were collected from C57BL/6 mice after 1 week (hypertrophy stage, HY) and 8 weeks post trans-aortic constriction (TAC) procedure (heart failure stage, HF), respectively, and their corresponding Sham controls (Sham-HY, Sham-HF). To conduct RNA-Seq analysis, total RNAs from six TAC and Sham-operated mice at the HY stage and four TAC and corresponding Sham mice at the HF stage were obtained.

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Transcriptome profiling of wildtype, nonoperated mouse hearts compared to wildtype, sham operated hearts

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