Characterization of fatty acid oxidation in human muscle mitochondria and myoblasts.
The mitochondrial oxidation of fatty acids (FAO) is the main energy-producing pathway in skeletal and cardiac muscle. Starting from standard muscle biopsies (100-200mg), we determined the optimal conditions of mitochondrial oxygen consumption by the FAO pathway, and in parallel we performed the isolation and primary culture of muscle cells to test their cellular FAO capacities. The determinations of maximal beta-oxidation rates in the presence of palmitoyl-CoA or palmitoyl-L-carnitine (mean+/-SEM: 32.5+/-2.0 and 34.1+/-1.3nmol O(2) min(-1) mg(-1) protein, n=16, respectively) provide a screening method of mitochondrial fatty acid transport system and intra-mitochondrial beta-oxidation. We also determined the conditions of tritiated palmitate oxidation by human myoblasts (mean+/-SEM: 6.6+/-0.1nmol 3H fatty acid h(-1) mg(-1) protein, n=8), and show that beta-oxidation defects can be detected in our experiments. Overall, we propose an original laboratory test to investigate FAO in human skeletal muscle and to screen for FAO disorders in myopathies and cardiomyopathies in human.
Djouadi
Fatima
F
INSERM Unité 393, Hôpital Necker--Enfants Malades, Paris, France.
Bonnefont
Jean Paul
JP
Munnich
Arnold
A
Bastin
Jean
J
D000293:Adolescent; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D006801:Humans; D008931:Mitochondria, Muscle / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D010084:Oxidation-Reduction; D019308:Palmitic Acid / Q000378:metabolism*
D019308:Palmitic Acid