UBC Theses and Dissertations
Respiratory properties of mitochondria from heart and mosaic muscle of rainbow trout (Salmo gairdneri) : substrate utilization and response to temperature and extramitochondrial pH Donaldson, Judith Margaret
Mitochondria were isolated from heart and mosaic muscle of rainbow trout (Salmo qairdneri R.). State 3 respiratory rates were determined at 5 and 15°C using pyruvate, malate, lactate, glutamate or acetyl—carnitine as substrate. The final three substrates were used to generate pH profiles. Pyruvate was oxidized at high rates in all cases, indicating good potential for aerobic carbohydrate metabolism. At 15°C, malate was an equally good substrate for heart mitochondria, while all substrates were oxidized at similar rates to pyruvate in muscle mitochondria. Maximal oxidation rates of heart mitochondria were greater than or equal to those of muscle. State 3 Q₁₀ for oxidation of most substrates in heart was approximately 2, except for malate which had a Q₁₀ of 3. Mitochondrial oxidation tended to be more sensitive to decreased temperature in muscle than in heart, particularly with respect to acetyl—carnitine and glutamate oxidation which in muscle had Q₁₀ values of 4 and 7, respectively. Based on RCR values at 5 and 15°C, there was no indication that membrane permeability to H⁺ ions was altered by a 10°C change in temperature in mitochondria from either tissue. At pH above 7.6 respiratory rates decreased with increasing pH. State 3 respiratory rate increased in heart mitochondria as pH decreased, below 7.6 while in muscle mitochondria, no such pH dependence was observed. RCR values were above 4 in all experiments except at high pH. Muscle mitochondria were the more sensitive to extreme pH with respect to RCR. Heart mitochondria had higher oxidative rates than those of muscle and were less sensitive to decreased temperature, in keeping with the greater oxidative demands of that tissue relative to mosaic muscle. Muscle mitochondria which typically face larger fluctuations in extramitochondrial pH in vivo than do those of heart, were less sensitive to pH in vitro. It was concluded that substrate utilization patterns and response to changes in temperature and extramitochondrial pH in the two mitochondrial populations was different and reflected both the intracellular environment of the mitochondria and the different needs of each tissue for aerobic energy supply.
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