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

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

Medicine

Producción científica: Contribución a una revista › Artículo › revisión exhaustiva

1 Cita (Scopus)

Resumen

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).

Idioma original	English
Publicación	STAR Protocols
DOI	https://doi.org/10.1016/j.xpro.2022.101738
Estado	Published - dic. 16 2022

Nota bibliográfica

Funding Information:
This research was supported by the Natural Sciences and Engineering Research Council of Canada , Research Nova Scotia , the Nova Scotia Research Innovation Trust and the Canada Foundation for Innovation . L.P.W. is supported by a Dalhousie Faculty of Medicine Scholarship . L.K.M., S.N., A.N., and E.B.C. are trainees in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute , with funds provided by the Dalhousie Medical Research Foundation (DMRF)’s Crease Endowment for Cancer Research to E.B.C. and funds provided by Craig’s Cause Pancreatic Cancer Society to A.N. L.K.M., S.N., and E.B.C. are recipients of Nova Scotia Graduate Scholarships. L.K.M. is additionally supported by a Genomics in Medicine Scholarship . S.N. and L.K.M. are additionally supported by a Killam Memorial Predoctoral Scholarship and the President’s award. S.N. is additionally funded by a CIHR Vanier award. A.N. is additionally funded by The Terry Fox Research Institute and Craig’s Cause Pancreatic Society. J.E.B. is the DMRF Cameron Cancer Scientist. The graphical abstract was created with biorender.com .

Funding Information:
This research was supported by the Natural Sciences and Engineering Research Council of Canada, Research Nova Scotia, the Nova Scotia Research Innovation Trust and the Canada Foundation for Innovation. L.P.W. is supported by a Dalhousie Faculty of Medicine Scholarship. L.K.M. S.N. A.N. and E.B.C. are trainees in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute, with funds provided by the Dalhousie Medical Research Foundation (DMRF)’s Crease Endowment for Cancer Research to E.B.C. and funds provided by Craig's Cause Pancreatic Cancer Society to A.N. L.K.M. S.N. and E.B.C. are recipients of Nova Scotia Graduate Scholarships. L.K.M. is additionally supported by a Genomics in Medicine Scholarship. S.N. and L.K.M. are additionally supported by a Killam Memorial Predoctoral Scholarship and the President's award. S.N. is additionally funded by a CIHR Vanier award. A.N. is additionally funded by The Terry Fox Research Institute and Craig's Cause Pancreatic Society. J.E.B. is the DMRF Cameron Cancer Scientist. The graphical abstract was created with biorender.com. We gratefully acknowledge the support of the Dalhousie Carlton Animal Care Facility. We are grateful to Dr. I.P. Alwayn for his for his assistance in initiating this project. Dalhousie University is located in Mi'kma’ki, the ancestral and unceded territory of the Mi'kmaq. Conceptualization, L.P.W. S.N. and J.E.B.; Methodology: L.P.W. and J.E.B.; Validation: L.P.W. S.N. A.N. L.K.M. E.B.C. and J.E.B.; Formal analysis: L.P.W. S.N. and J.E.B.; Investigation: L.P.W. S.N. A.N. L.K.M. and E.B.C.; Data curation: L.P.W. L.K.M. and S.N.; Writing-original draft: L.P.W. and J.E.B.; Writing Review & Editing: L.P.W. S.N. A.N. L.K.M. E.B.C. and J.E.B.; Visualization: L.P.W. S.N. and J.E.B. Supervision: J.E.B.; Project administration: L.P.W. S.N. and J.E.B.; Funding acquisition: J.E.B. The authors declare no competing interests.

Publisher Copyright:
© 2022 The Author(s)

ASJC Scopus Subject Areas

General Neuroscience
General Biochemistry,Genetics and Molecular Biology
General Immunology and Microbiology

PubMed: MeSH publication types

Journal Article

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10.1016/j.xpro.2022.101738

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@article{916323b8b0d94713b19eef4c05ad0cd4,

title = "Preparation of mitochondrial damage-associated molecular patterns from mouse liver tissue",

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).",

author = "Westhaver, {Lauren P.} and Sarah Nersesian and Adam Nelson and MacLean, {Leah K.} and Carter, {Emily B.} and Boudreau, {Jeanette E.}",

note = "Funding Information: This research was supported by the Natural Sciences and Engineering Research Council of Canada , Research Nova Scotia , the Nova Scotia Research Innovation Trust and the Canada Foundation for Innovation . L.P.W. is supported by a Dalhousie Faculty of Medicine Scholarship . L.K.M., S.N., A.N., and E.B.C. are trainees in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute , with funds provided by the Dalhousie Medical Research Foundation (DMRF){\textquoteright}s Crease Endowment for Cancer Research to E.B.C. and funds provided by Craig{\textquoteright}s Cause Pancreatic Cancer Society to A.N. L.K.M., S.N., and E.B.C. are recipients of Nova Scotia Graduate Scholarships. L.K.M. is additionally supported by a Genomics in Medicine Scholarship . S.N. and L.K.M. are additionally supported by a Killam Memorial Predoctoral Scholarship and the President{\textquoteright}s award. S.N. is additionally funded by a CIHR Vanier award. A.N. is additionally funded by The Terry Fox Research Institute and Craig{\textquoteright}s Cause Pancreatic Society. J.E.B. is the DMRF Cameron Cancer Scientist. The graphical abstract was created with biorender.com . Funding Information: This research was supported by the Natural Sciences and Engineering Research Council of Canada, Research Nova Scotia, the Nova Scotia Research Innovation Trust and the Canada Foundation for Innovation. L.P.W. is supported by a Dalhousie Faculty of Medicine Scholarship. L.K.M. S.N. A.N. and E.B.C. are trainees in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute, with funds provided by the Dalhousie Medical Research Foundation (DMRF){\textquoteright}s Crease Endowment for Cancer Research to E.B.C. and funds provided by Craig's Cause Pancreatic Cancer Society to A.N. L.K.M. S.N. and E.B.C. are recipients of Nova Scotia Graduate Scholarships. L.K.M. is additionally supported by a Genomics in Medicine Scholarship. S.N. and L.K.M. are additionally supported by a Killam Memorial Predoctoral Scholarship and the President's award. S.N. is additionally funded by a CIHR Vanier award. A.N. is additionally funded by The Terry Fox Research Institute and Craig's Cause Pancreatic Society. J.E.B. is the DMRF Cameron Cancer Scientist. The graphical abstract was created with biorender.com. We gratefully acknowledge the support of the Dalhousie Carlton Animal Care Facility. We are grateful to Dr. I.P. Alwayn for his for his assistance in initiating this project. Dalhousie University is located in Mi'kma{\textquoteright}ki, the ancestral and unceded territory of the Mi'kmaq. Conceptualization, L.P.W. S.N. and J.E.B.; Methodology: L.P.W. and J.E.B.; Validation: L.P.W. S.N. A.N. L.K.M. E.B.C. and J.E.B.; Formal analysis: L.P.W. S.N. and J.E.B.; Investigation: L.P.W. S.N. A.N. L.K.M. and E.B.C.; Data curation: L.P.W. L.K.M. and S.N.; Writing-original draft: L.P.W. and J.E.B.; Writing Review & Editing: L.P.W. S.N. A.N. L.K.M. E.B.C. and J.E.B.; Visualization: L.P.W. S.N. and J.E.B. Supervision: J.E.B.; Project administration: L.P.W. S.N. and J.E.B.; Funding acquisition: J.E.B. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = dec,

day = "16",

doi = "10.1016/j.xpro.2022.101738",

language = "English",

journal = "STAR Protocols",

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publisher = "Cell Press",

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TY - JOUR

T1 - Preparation of mitochondrial damage-associated molecular patterns from mouse liver tissue

AU - Westhaver, Lauren P.

AU - Nersesian, Sarah

AU - Nelson, Adam

AU - MacLean, Leah K.

AU - Carter, Emily B.

AU - Boudreau, Jeanette E.

N1 - Funding Information: This research was supported by the Natural Sciences and Engineering Research Council of Canada , Research Nova Scotia , the Nova Scotia Research Innovation Trust and the Canada Foundation for Innovation . L.P.W. is supported by a Dalhousie Faculty of Medicine Scholarship . L.K.M., S.N., A.N., and E.B.C. are trainees in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute , with funds provided by the Dalhousie Medical Research Foundation (DMRF)’s Crease Endowment for Cancer Research to E.B.C. and funds provided by Craig’s Cause Pancreatic Cancer Society to A.N. L.K.M., S.N., and E.B.C. are recipients of Nova Scotia Graduate Scholarships. L.K.M. is additionally supported by a Genomics in Medicine Scholarship . S.N. and L.K.M. are additionally supported by a Killam Memorial Predoctoral Scholarship and the President’s award. S.N. is additionally funded by a CIHR Vanier award. A.N. is additionally funded by The Terry Fox Research Institute and Craig’s Cause Pancreatic Society. J.E.B. is the DMRF Cameron Cancer Scientist. The graphical abstract was created with biorender.com . Funding Information: This research was supported by the Natural Sciences and Engineering Research Council of Canada, Research Nova Scotia, the Nova Scotia Research Innovation Trust and the Canada Foundation for Innovation. L.P.W. is supported by a Dalhousie Faculty of Medicine Scholarship. L.K.M. S.N. A.N. and E.B.C. are trainees in the Cancer Research Training Program of the Beatrice Hunter Cancer Research Institute, with funds provided by the Dalhousie Medical Research Foundation (DMRF)’s Crease Endowment for Cancer Research to E.B.C. and funds provided by Craig's Cause Pancreatic Cancer Society to A.N. L.K.M. S.N. and E.B.C. are recipients of Nova Scotia Graduate Scholarships. L.K.M. is additionally supported by a Genomics in Medicine Scholarship. S.N. and L.K.M. are additionally supported by a Killam Memorial Predoctoral Scholarship and the President's award. S.N. is additionally funded by a CIHR Vanier award. A.N. is additionally funded by The Terry Fox Research Institute and Craig's Cause Pancreatic Society. J.E.B. is the DMRF Cameron Cancer Scientist. The graphical abstract was created with biorender.com. We gratefully acknowledge the support of the Dalhousie Carlton Animal Care Facility. We are grateful to Dr. I.P. Alwayn for his for his assistance in initiating this project. Dalhousie University is located in Mi'kma’ki, the ancestral and unceded territory of the Mi'kmaq. Conceptualization, L.P.W. S.N. and J.E.B.; Methodology: L.P.W. and J.E.B.; Validation: L.P.W. S.N. A.N. L.K.M. E.B.C. and J.E.B.; Formal analysis: L.P.W. S.N. and J.E.B.; Investigation: L.P.W. S.N. A.N. L.K.M. and E.B.C.; Data curation: L.P.W. L.K.M. and S.N.; Writing-original draft: L.P.W. and J.E.B.; Writing Review & Editing: L.P.W. S.N. A.N. L.K.M. E.B.C. and J.E.B.; Visualization: L.P.W. S.N. and J.E.B. Supervision: J.E.B.; Project administration: L.P.W. S.N. and J.E.B.; Funding acquisition: J.E.B. The authors declare no competing interests. Publisher Copyright: © 2022 The Author(s)

PY - 2022/12/16

Y1 - 2022/12/16

N2 - 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).

AB - 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).

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

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ASJC Scopus Subject Areas

PubMed: MeSH publication types

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