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A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation
Published on: July 13, 2018
Cardiac myocyte gene expression profiling during H2O2-induced apoptosis
Angela Clerk1, Timothy J Kemp, Georgia Zoumpoulidou
1National Heart and Lung Institute Division, Faculty of Medicine, Imperial College London, London, United Kingdom. a.clerk@imperial.ac.uk
Physiological Genomics
|December 7, 2006
Summary
High hydrogen peroxide (H2O2) levels induce cardiac myocyte death. This study identified novel H2O2-responsive genes and dynamic gene expression changes during apoptosis in rat heart cells.
Area of Science:
- Cardiovascular Biology
- Molecular Biology
- Oxidative Stress Research
Background:
- Oxidative stress influences cardiac myocyte survival, with high levels promoting death and lower levels potentially being cytoprotective.
- Hydrogen peroxide (H2O2) is a key mediator of oxidative stress.
Purpose of the Study:
- To investigate gene expression changes in rat neonatal cardiac myocytes exposed to varying concentrations of H2O2.
- To identify novel genes responsive to H2O2 and understand their role in apoptosis.
Main Methods:
- Affymetrix microarrays (U34B and 230 2.0) were used to analyze gene expression profiles.
- Rat neonatal cardiac myocytes were treated with apoptotic (0.2 mM) and non-toxic (0.04 mM) H2O2 concentrations for 2, 4, or 24 hours.
- Quantitative PCR was employed to validate specific gene expression changes, including Krüppel-like factors (Klfs).
Main Results:
- A novel H2O2-responsive gene, putative peroxide-inducible transcript 1 (Perit1), generating two alternatively spliced transcripts, was identified.
- Low H2O2 concentrations (<0.1 mM) did not increase protein synthesis, indicating they are not globally anabolic in this system.
- Apoptotic H2O2 (0.2 mM) significantly altered the expression of 649 mRNAs and 187 uncharacterized RNAs, including 114 transcriptional regulators like Klfs.
Conclusions:
- Hydrogen peroxide-induced cardiac myocyte apoptosis is characterized by significant and dynamic alterations in gene expression.
- The identified genes and their protein products may play a crucial role in modulating the apoptotic response in cardiac myocytes.
