Isolation and functional analysis of peridroplet mitochondria from murine brown adipose tissue

Jennifer Ngo; Ilan Y Benador; Alexandra J Brownstein; Laurent Vergnes; Michaela Veliova; Michael Shum; Rebeca Acín-Pérez; Karen Reue; Orian S Shirihai; Marc Liesa

doi:10.1016/j.xpro.2020.100243

Isolation and functional analysis of peridroplet mitochondria from murine brown adipose tissue

STAR Protoc. 2020 Dec 30;2(1):100243. doi: 10.1016/j.xpro.2020.100243. eCollection 2021 Mar 19.

Authors

Jennifer Ngo^{1

2

3}, Ilan Y Benador¹, Alexandra J Brownstein^{1

3}, Laurent Vergnes^{3

4}, Michaela Veliova^{1

3}, Michael Shum^{1

3}, Rebeca Acín-Pérez^{1

3}, Karen Reue^{3

4

5}, Orian S Shirihai^{1

3}, Marc Liesa^{1

3

5}

Affiliations

¹ Department of Medicine, Division of Endocrinology and Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
² Department of Chemistry and Biochemistry at UCLA, Los Angeles, CA 90095, USA.
³ Metabolism Theme, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095, USA.
⁴ Department of Human Genetics, David Geffen School of Medicine at UCLA, Los Angeles, CA 90024, USA.
⁵ Molecular Biology Institute at UCLA, Los Angeles, CA 90095, USA.

Abstract

Mitochondria play a central role in lipid metabolism and can bind to lipid droplets. However, the role and functional specialization of the population of peridroplet mitochondria (PDMs) remain unclear, as methods to isolate functional PDMs were not developed until recently. Here, we describe an approach to isolate intact PDMs from murine brown adipose tissue based on their adherence to lipid droplets. PDMs isolated using our approach can be used to study their specialized function by respirometry. For complete information on the use and execution of this protocol, please refer to Benador et al. (2018).

Keywords: Metabolism; Molecular biology.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adipocytes, Brown / cytology
Adipocytes, Brown / metabolism*
Adipose Tissue, Brown / cytology
Adipose Tissue, Brown / metabolism*
Animals
Lipid Droplets / metabolism*
Lipid Metabolism*
Mice
Mitochondria / metabolism*

Abstract

Publication types

MeSH terms

Grants and funding