New insights into the regulation of plant succinate dehydrogenase. On the role of the protonmotive force.

Regulation of succinate dehydrogenase was investigated using tightly coupled potato tuber mitochondria in a novel fashion by simultaneously measuring the oxygen uptake rate and the ubiquinone (Q) reduction level. We found that the activation level of the enzyme is unambiguously reflected by the kinetic dependence of the succinate oxidation rate upon the Q-redox poise. Kinetic results indicated that succinate dehydrogenase is activated by both ATP (K(1/2) approximately 3 microm) and ADP. The carboxyatractyloside insensitivity of these stimulatory effects indicated that they occur at the cytoplasmic side of the mitochondrial inner membrane. Importantly, our novel approach revealed that the enzyme is also activated by oligomycin (K(1/2) approximately 16 nm). Time-resolved kinetic measurements of succinate dehydrogenase activation by succinate furthermore revealed that the activity of the enzyme is negatively affected by potassium. The succinate-induced activation (+/-K(+)) is prevented by the presence of an uncoupler. Together these results demonstrate that in vitro activity of succinate dehydrogenase is modulated by the protonmotive force. We speculate that the widely recognized activation of the enzyme by adenine nucleotides in plants is mediated in this manner. A mechanism that could account for such regulation is suggested and ramifications for its in vivo relevance are discussed.

Kinetic analysis of the mitochondrial quinol-oxidizing enzymes during development of thermogenesis in Arum maculatum L.

The dependence of the rate of oxygen uptake upon the ubiquinone (Q)-pool reduction level in mitochondria isolated during the development of thermogenesis of Arum maculatum spadices has been investigated. At the alpha-stage of development, the respiratory rate was linearly dependent upon the reduction level of the Q-pool (Qr) both under state-3 and -4 conditions. Progression through the beta/gamma to the delta-stage resulted in a non-linear dependence of the state-4 rate on Qr. In the delta-stage of development, both state-3 and -4 respiratory rates were linearly dependent upon Qr due to a shift in the engagement of the alternative oxidase to lower levels of Qr. Western blot analysis revealed that increased alternative oxidase activity could be correlated with expression of a 35 kDa protein. Respiratory control was only observed with mitochondria in the alpha-stage of development. At the beta/gamma-stage of development, the addition of ADP resulted in a significant oxidation of the Q-pool which was accompanied by a decrease in the respiratory rate. This was due either to decreased contribution of the alternative pathway to the overall respiratory rate under state 3 or by deactivation of succinate dehydrogenase activity by ADP. Cold-storage of the spadices at the beta-stage of development led to increased activity of both the cytochrome pathway and succinate dehydrogenase, without any change in alternative oxidase activity. Results are discussed in terms of how changes in the activation level of the alternative oxidase and succinate dehydrogenase influence the activity and engagement of the quinol-oxidizing pathways during the development of thermogenesis in A. maculatum.

Growth of broiler chickens in response to feed restriction regimens to reduce ascites.

A trial was conducted to investigate the impact of early feed restriction on ascites induced by cold temperatures and the subsequent effect on the whole body and breast muscle growth of broilers. Two feed restriction regimens were tested, consisting of limiting daily feed intake of the birds to 75% of the ME required for normal growth from either 4 to 11 d or from 7 to 14 d. At 21 d, half of the birds were moved to a cold house (17.8 C) to induce ascites. Five birds from each pen were killed for the breast muscle growth, Pectoralis major and Pectoralis minor, and heart and abdominal fat pad weights at 4, 7, 11, 14, 21, 35, and 49 d of the experiment. Birds in the cold house were heavier and had better feed conversion than birds in the control house at 49 d of age. This could be attributed to the high ambient temperatures (27 to 33 C) in the control house. The ad libitum birds had a significantly greater percentage of P. major than the feed-restricted birds. Exposure to cold temperatures caused significantly higher percentage of ascites from 21 to 49 d. Cold temperatures also increased the percentage of total heart and the right ventricle weight relative to total heart weight ratio at 35 and 49 d of age, suggesting that the surviving birds were more likely to develop ascites. Although catch-up growth was observed, the final body weight of feed-restricted birds was not the same as ad libitum birds. Early feed restriction reduced the incidence of ascites, but at the cost of breast muscle growth.