Isolation of mitochondrial subpopulations from skeletal muscle: Optimizing recovery and preserving integrity.

ABSTRACT:

Aim

The subsarcolemmal (SSM) and interfibrillar (IFM) mitochondria in skeletal muscle appear to have distinct biochemical properties affecting metabolism in health and disease. The isolation of mitochondrial subpopulations has been a long-time challenge while the presence of a continuous mitochondrial reticulum challenges the view of distinctive SSM and IFM bioenergetics. Here, a comprehensive approach is developed to identify the best conditions to separate mitochondrial fractions.

Methods

The main modifications to the protocol to isolate SSM and IFM from rat skeletal muscle were: (a) decreased dispase content and homogenization speed; (b) trypsin treatment of SSM fractions; (c) recentrifugation of mitochondrial fractions at low speed to remove subcellular components. To id

SUBMITTER: Lai N 

PROVIDER: S-EPMC6335164 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature