Simultaneous preparation of membrane fractions from small amounts of skeletal muscle: a study on mitochondrial and microsomal fractions from MedJ mice.
This work is the first biochemical study of skeletal muscle membranes isolated from mice displaying an inherited neuromuscular disease: MedJ strains. It is focused on the research of a possible alteration of membrane biological activities related to this disease. We describe a procedure which allows the simultaneous preparation of mitochondrial and microsomal fractions from a small amount of skeletal muscle. When EGTA and BSA are present in the buffers, functional mitochondria can be prepared. Under these conditions we found that no major modification occurs for this disease at the mitochondrial inner membrane level. A dramatic impairment of a calcium active transport activity found in the microsomal fraction obtained from MedJ is noticed, suggesting that some modification may occur at this level.
Rock
E
E
Napias
C
C
Sarger
C
C
Chevallier
J
J
D000818:Animals; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D002458:Cell Fractionation; D007425:Intracellular Membranes / Q000648:ultrastructure*; D051379:Mice; D008817:Mice, Mutant Strains; D008861:Microsomes / Q000648:ultrastructure; D008931:Mitochondria, Muscle / Q000648:ultrastructure; D009132:Muscles / Q000201:enzymology / Q000648:ultrastructure*; D009468:Neuromuscular Diseases / Q000473:pathology*; D010101:Oxygen Consumption
D000252:Calcium-Transporting ATPases; D002118:Calcium