Preparation of mitochondrial damage-associated molecular patterns from mouse liver tissue

Lauren P Westhaver; Sarah Nersesian; Adam Nelson; Leah K MacLean; Emily B Carter; Jeanette E Boudreau

doi:10.1016/j.xpro.2022.101738

Preparation of mitochondrial damage-associated molecular patterns from mouse liver tissue

STAR Protoc. 2022 Dec 16;3(4):101738. doi: 10.1016/j.xpro.2022.101738. Epub 2022 Sep 30.

Authors

Lauren P Westhaver¹, Sarah Nersesian², Adam Nelson², Leah K MacLean³, Emily B Carter², Jeanette E Boudreau⁴

Affiliations

¹ Department of Pathology, Dalhousie University, Sir Charles Tupper Medical Building, 5850 College St., Halifax, NS B3H 4H7, Canada. Electronic address: lauren.westhaver@dal.ca.
² Department of Microbiology and Immunology, Dalhousie University, Room 7C, Sir Charles Tupper Medical Building, 5850 College St., Halifax, NS B3H 4H7, Canada; Beatrice Hunter Cancer Research Institute, Halifax, NS, Canada.
³ Department of Pathology, Dalhousie University, Sir Charles Tupper Medical Building, 5850 College St., Halifax, NS B3H 4H7, Canada; Beatrice Hunter Cancer Research Institute, Halifax, NS, Canada.
⁴ Department of Pathology, Dalhousie University, Sir Charles Tupper Medical Building, 5850 College St., Halifax, NS B3H 4H7, Canada; Department of Microbiology and Immunology, Dalhousie University, Room 7C, Sir Charles Tupper Medical Building, 5850 College St., Halifax, NS B3H 4H7, Canada; Beatrice Hunter Cancer Research Institute, Halifax, NS, Canada. Electronic address: jeanette.boudreau@dal.ca.

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).

Keywords: Cell Biology; Cell isolation; Cell separation/fractionation; Cell-based Assays; Immunology; Metabolomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Liver*
Mice
Mitochondria*

Grants and funding

CIHR/Canada