Preparation of mitochondrial damage-associated molecular patterns from mouse liver tissue
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Published on: November 22, 2017
Lauren P Westhaver1, Sarah Nersesian2, Adam Nelson2
1Department of Pathology, Dalhousie University, Sir Charles Tupper Medical Building, 5850 College St., Halifax, NS B3H 4H7, Canada.
Abstract:
Mitochondrial damage-associated molecular patterns (mitoDAMPs) are released from cells dying uncontrolled, non-apoptotic deaths, usually secondary to disease or trauma. Here, we describe preparation of mitoDAMPs from mouse liver, but this protocol can be adapted for preparation of mitoDAMPs from other species and tissues. Tissues are dissociated and then processed to isolate mitochondria. Mitochondria are then sonicated and mitoDAMPs are collected by ultracentrifugation. This procedure produces μg quantities of mitoDAMPs and facilitates research to understand their impacts in health and disease. For complete details on the use and execution of this protocol, please refer to Westhaver et al. (2022).
Insights
Researchers developed a protocol to isolate mitochondrial damage-associated molecular patterns (mitoDAMPs) from mouse liver. This method aids in studying the role of mitoDAMPs in various health and disease conditions.
Area of Science:
- Biochemistry
- Immunology
- Cell Biology
Background:
- Mitochondrial damage-associated molecular patterns (mitoDAMPs) are released during uncontrolled cell death.
- These molecules play a role in inflammatory responses following disease or trauma.
Purpose of the Study:
- To describe a standardized protocol for the preparation of mitoDAMPs.
- To facilitate research on the biological functions of mitoDAMPs.
Main Methods:
- Isolation of mitochondria from mouse liver tissue.
- Sonication of isolated mitochondria to release mitoDAMPs.
- Collection of mitoDAMPs via ultracentrifugation.
Main Results:
- Successful preparation of microgram quantities of mitoDAMPs.
- The protocol is adaptable to different species and tissues.
Conclusions:
- This protocol provides a reliable method for mitoDAMPs preparation.
- Enables further investigation into mitoDAMPs' roles in physiological and pathological processes.
