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Updated: Apr 6, 2026
Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue
Published on: March 23, 2015
Analysis of cytosolic mtDNA release during Staphylococcus aureus infection
Caterina Licini1, Gloria D'Achille2, Nada Dhaouadi1
1Department of Clinical and Molecular Sciences, Marche Polytechnic University, Ancona, Italy.
Abstract:
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the principal human pathogens, causing severe infections in skin wounds. MRSA infection triggers a cell response mainly by mitochondrial-mediated pathway, resulting in mitochondrial outer membrane permeabilization, extrusion of the mitochondrial inner membrane into the cytoplasm, and then spillage of mitochondrial DNA (mtDNA) into the cytoplasm. The cell recognizes the discharged cytosolic mtDNA (cmtDNA) as "not-itself" because of mtDNA properties and triggers cascade events, such as the activation of inflammasomes. Here, we detail a method to detect and measure the mtDNA release into the cytoplasm in immortalized keratinocytes (HaCaT cells), after the infection with MRSA at different time points after the infection.
Insights
Methicillin-resistant Staphylococcus aureus (MRSA) infection releases mitochondrial DNA (mtDNA) into the cytoplasm of skin cells. This study details a method to detect and measure this critical mtDNA release following MRSA infection.
Area of Science:
- Cell Biology
- Microbiology
- Immunology
Background:
- Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen causing severe skin wound infections.
- MRSA infection initiates a cellular response via mitochondrial pathways, leading to mitochondrial DNA (mtDNA) release into the cytoplasm.
- Cytosolic mtDNA (cmtDNA) is recognized as foreign, triggering inflammatory responses like inflammasome activation.
Purpose of the Study:
- To detail a method for detecting and quantifying mtDNA release into the cytoplasm.
- To investigate mtDNA release in immortalized keratinocytes (HaCaT cells) after MRSA infection.
- To analyze mtDNA release kinetics at various time points post-infection.
Main Methods:
- Utilized immortalized keratinocytes (HaCaT cells).
- Infected cells with Methicillin-resistant Staphylococcus aureus (MRSA).
- Developed and applied a method to detect and measure cytosolic mtDNA release over time.
Main Results:
- Successfully detected and measured the release of mtDNA into the cytoplasm of HaCaT cells post-MRSA infection.
- Quantified mtDNA release at different time intervals following infection.
- Established a quantifiable assay for studying MRSA-induced mtDNA release.
Conclusions:
- The study provides a reliable method to assess mtDNA release during MRSA infection in keratinocytes.
- Understanding mtDNA release is crucial for elucidating inflammatory responses to bacterial pathogens.
- This method can be applied to study host-pathogen interactions and develop targeted therapies.
