Analysis of age-associated changes in mitochondrial free radical generation by rat testis.

Throughout spermatogenesis, mitochondria undergo a morphological and functional differentiation. Mitochondria are involved in the production of reactive oxygen species (ROS), considered one of the mediators of ageing. Particularly, lipid peroxidation is regarded as a major phenomenon by which ROS can impair cellular function. In the present study, we examined the production of superoxide anion, superoxide dismutase activity and the effect of Fe(2+)/ascorbate induced-lipid peroxidation on the respiratory chain activities of testis mitochondria throughout the process of spermatogenesis and ageing. Mitochondria from rat testes generated superoxide anion, mainly using NADH as substrate, which increased according to age. The activity of SOD is age-dependent and greatly stimulated during the first wave of spermatogenesis, but decreases in adulthood and old age. TBARS concentration was also markedly increased by ageing. The activity of mitochondrial respiratory chain complexes is differentially affected by oxidative stress induced by iron/ascorbate, succinate-dehydrogenase activity being less vulnerable than that of NADH-dehydrogenase and cytochrome c oxidase. The data suggest that ageing is accompanied by reduced activity of SOD, leading to excessive oxidative stress and enhanced lipid peroxidation that compromises the functionality of the electron transport chain. The data support the concept that mitochondrial function is an important determinant in ageing.

Respiratory chain complexes and membrane fatty acids composition in rat testis mitochondria throughout development and ageing.

Throughout the maturation of germ cells, a morphological, biochemical and functional differentiation of mitochondria has been shown to occur. Ageing is known to cause changes involved in energy metabolism. These changes have been related to molecular and functional alterations in the properties of biological membranes. Variations in membrane lipid composition and lipid-protein interactions occur with ageing in several tissues. The present paper describes the relationship between these membrane alterations and the activities of lipid-dependent enzymes of isolated testis mitochondria in rats of from 10 days of age to 24 months. The specific activities of these enzymes are lower in preparations from adult and aged rats as compared to those from young rats. Temperature breaks of Arrhenius plots show age-dependent shifts to higher temperatures for the NADH-dehydrogenase, succinate-dehydrogenase, cytochrome c oxidase, and ATPase in senescent animals. Analysis of the membrane fatty acid composition reveals a distinct age-dependent fall in the content of polyunsaturated fatty acids accompanied by an increase in the proportion of saturated fatty acids and a decrease in polyunsaturated fatty acid percentage. The results suggest that during spermatogenesis and the ageing process some changes in the composition of the fatty acids in the surrounding membrane affect the protein-lipid interactions, producing a decrease in mitochondrial enzyme activities.