Different yield and properties of mitochondria from skeletal muscle of normal hamsters

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Abstract
Yield and properties of mitochondria, isolated from hind leg muscles of normal hamsters, are different, when 4 preparation procedures are applied simultaneously: 1. mechanical homogenization of muscle with a glass-Teflon homogenizer followed by rehomogenization of the isolated nuclear fraction for 1–6 times. The procedure yields impure mitochondria showing the highest amounts of mitochondrial enzymes (50–65% of total), protein and DNA; the isolated mitochondria exhibit disrupted outer membranes and loosened inner membranes causing a better supply with substrates of enzymes located at the inner membrane and in the matrix. 2. Single mechanical homogenization of muscle for increasing periods (2–40 min) yields lower amounts of all constituents (25–40% of total). 3. The yields of Nagarse treatment with one step of 2 min mechanical homogenization range between 20–35% of total. Mitochondria isolated with the latter procedure exhibit compact membranes and are very pure, but still contaminated with Nagarse, causing destructions of mitochondrial membranes already after 24 h storage in buffer with sucrose and mannitol at 2°C. 4. One single Nagarse treatment of muscle for 40 min produces a minor yield of these constituents. Therefore, at least two different isolation procedures are proposed for optimum investigation of skeletal muscle mitochondria.