Temperature and sex dependent effects on cardiac mitochondrial metabolism in Atlantic cod (Gadus morhua L.).

To test the hypothesis that impaired mitochondrial respiration limits cardiac performance at warm temperatures, and examine if any effect(s) are sex-related, the consequences of high temperature on cardiac mitochondrial oxidative function were examined in 10°C acclimated, sexually immature, male and female Atlantic cod. Active (State 3) and uncoupled (States 2 and 4) respiration were measured in isolated ventricular mitochondria at 10, 16, 20, and 24°C using saturating concentrations of malate and pyruvate, but at a submaximal (physiological) level of ADP (200µM). In addition, citrate synthase (CS) activity was measured at these temperatures, and mitochondrial respiration and the efficiency of oxidative phosphorylation (P:O ratio) were determined at [ADP] ranging from 25-200µM at 10 and 20°C. Cardiac morphometrics and mitochondrial respiration at 10°C, and the thermal sensitivity of CS activity (Q10=1.51), were all similar between the sexes. State 3 respiration at 200µM ADP increased gradually in mitochondria from females between 10 and 24°C (Q10=1.48), but plateaued in males above 16°C, and this resulted in lower values in males vs. females at 20 and 24°C. At 10°C, State 4 was ~10% of State 3 values in both sexes [i.e. a respiratory control ratio (RCR) of ~10] and P:O ratios were approximately 1.5. Between 20 and 24°C, State 4 increased more than State 3 (by ~70 vs. 14%, respectively), and this decreased RCR to ~7.5. The P:O ratio was not affected by temperature at 200μM ADP. However, (1) the sensitivity of State 3 respiration to increasing [ADP] (from 25 to 200μM) was reduced at 20 vs. 10°C in both sexes (Km values 105±7 vs. 68±10μM, respectively); and (2) mitochondria from females had lower P:O values at 25 vs. 100μM ADP at 20°C, whereas males showed a similar effect at 10°C but a much more pronounced effect at 20°C (P:O 1.05 at 25μM ADP vs. 1.78 at 100μMADP). In summary, our results demonstrate several sex-related differences in ventricular mitochondrial function in Atlantic cod, and suggest that myocardial oxidative function and possibly phosphorylation efficiency may be limited at temperatures of 20°C or above, particularly in males. These observations could partially explain why cardiac function in Atlantic cod plateaus just below this species׳ critical thermal maximum (~22°C) and may contribute to yet unidentified sex differences in thermal tolerance and swimming performance.