Mitochondrial Isolation and Purification from Mouse Spinal Cord

Abstract

Mitochondria are eukaryotic organelles that play a crucial role in several cellular processes, including energy production, β-oxidation of fatty acids and regulation of calcium homeostasis. In the last 20 years there has been a hightened interest in the study of mitochondria following the discoveries that mitochondria are central to the process of programmed cell death and that mitochondrial dysfunctions are implicated in numerous diseases including a wide range of neurological disorders such as Parkinson’s disease, Alzheimer’s disease, Huntington’s disease and amyotrophic lateral sclerosis. In order to identify and study changes in mitochondrial function related to specific neurological conditions the mitochondria are often isolated from the compartment of the central nervous system most affected during disease. Here, we describe a protocol for the isolation of mitochondria from mouse spinal cord, a compartment of the central nervous system that is significantly affected in neuromuscular diseases such as amyotrophic lateral sclerosis. This method relies on differential centrifugation to separate the mitochondria from the other subcellular compartments.