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Updated: Oct 22, 2025
Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue
Published on: March 23, 2015
Isolation and analysis of fractions enriched in WrappER-associated mitochondria from mouse liver
Nicolò Ilacqua1,2, Irene Anastasia1,2, Luca Pellegrini2,3
1Graduate Program in Neuroscience, Faculty of Medicine, Laval University, Quebec, QC, Canada.
Abstract:
The endoplasmic reticulum (ER) plays a central role in lipid homeostasis, but the role of individual ER subdomains in lipid biology has not been elucidated. WrappER is a curved wrapping type of rough-ER that establishes extensive contacts with almost every mitochondria of the hepatocyte in the mouse liver. Here, we describe a protocol for isolation of fractions enriched in wrappER-associated mitochondria from the mouse liver. We also provide techniques for assessing its quality by electron microscopy and biochemical/proteomic analysis. For complete information on the use and execution of this protocol, please refer to Anastasia et al. (2021).
Insights
This study details a protocol for isolating specialized endoplasmic reticulum (ER) structures called WrappER, which are closely associated with mitochondria in mouse liver cells. This method aids in studying ER-mitochondria interactions and lipid homeostasis.
Area of Science:
- Cell Biology
- Mitochondrial Biology
- Lipid Metabolism
Background:
- The endoplasmic reticulum (ER) is crucial for lipid homeostasis.
- Specific roles of ER subdomains in lipid biology remain unclear.
- WrappER, a unique rough-ER subdomain, extensively contacts mitochondria in hepatocytes.
Purpose of the Study:
- To describe a protocol for isolating WrappER-associated mitochondria from mouse liver.
- To provide methods for quality assessment of isolated fractions.
- To facilitate research into ER-mitochondria interactions and lipid biology.
Main Methods:
- Isolation of fractions enriched in WrappER-associated mitochondria from mouse liver.
- Quality assessment using electron microscopy.
- Biochemical and proteomic analysis for fraction characterization.
Main Results:
- Successful isolation protocol for WrappER-associated mitochondria.
- Established methods for verifying the purity and integrity of isolated fractions.
- Provides a foundation for further investigation into the function of these specific ER-mitochondria contacts.
Conclusions:
- The described protocol enables the study of specialized ER subdomains (WrappER) and their mitochondrial interactions.
- This method is valuable for understanding the role of ER-mitochondria contacts in hepatic lipid metabolism.
- Further research can utilize this protocol to explore the functional significance of WrappER-mitochondria associations.
