Loose-coupled subsarcolemmal mitochondria from muscle Rhomboideus in cold-acclimated piglets

1. Intermyofibrillar (IM) and subsarcolemmal (SM) mitochondria were isolated from rhomboideus (RH) and longissimus dorsi (LD) muscles of cold-acclimated (12 degrees C for 3 weeks) and control (23 degrees C) 8-week-old piglets. 2. Together with measurements of yield of mitochondrial protein and enzyme activities (cytochrome oxydase-CO; creatine kinase--CK), the respiratory rate of isolated mitochondria was followed polarographically in order to determine the respiratory control ratio (RCR) and consequently the tightness of coupling in response to ADP. 3. In control and cold-acclimated piglets, there were more IM than SM (P less than 0.05) and more mitochondria in RH than LD muscle (P less than 0.05). In both muscles, the yield of mitochondria was slightly but not significantly higher after cold acclimation than in controls. 4. In both muscles, IM were tightly coupled and their RCR (congruent to 4.5) were similar in both groups of piglets. RCR values were increased in the presence of bovine serum albumin (BSA). 5. In controls, SM exhibited lower respiration rates than IM (P less than 0.05) and were slightly coupled (RCR congruent to 2). Cold acclimation increases the loose-coupling of SM (P less than 0.05), especially in RH muscle. No changes appeared in the mitochondrial coupling after the addition of BSA. 6. After cold acclimation, CO and CK activities were increased in IM (P less than 0.05) while only CO activity was increased in SM (P less than 0.05). These results support a coupling defect in SM and therefore confirm mitochondrial respiration results.