Hiren R. Modi

Ageing-induced alterations in lipid/phospholipid profiles of rat brain and liver mitochondria: implications for mitochondrial energy-linked functions.

Effects of ageing on the lipid/phospholipid profile of brain and liver mitochondria from rats were examined. In the brain mitochondria the contents of total phospholipid (TPL) and cholesterol (CHL) increased with simultaneous increase in the TPL/CHL (mole:mole) ratio. The proportion and contents of lysophospholipid (Lyso), sphingomyelin (SPM), phosphatidylinositol (PI), phosphatidylserine (PS) and diphosphatidylglycerol (DPG) components increased, with maximal increases seen for PS and PI; phosphatidylcholine (PC) and phosphatidylethanolamine (PE) components registered decrease. In the liver mitochondria contents of TPL and CHL increased. However, the TPL/CHL (mole:mole) ratio was not altered. Lyso, PI and PS increased. However, the magnitude of increase was competitively lower; PE and DPG decreased. SPM and PC did not change as a consequence of ageing. These changes altered the contents of individual phospholipids in the two membrane systems. Respiration with glutamate, pyruvate + malate, succinate and ascorbate + N,N,N’,N’-tetramethyl-p-phenylenediamine was significantly impaired in brain mitochondria from old animals. For liver mitochondria the respiratory activity declined with glutamate and succinate. Correlation studies by regression analysis revealed that the lipid/phospholipid classes regulate respiratory function differently in the mitochondria from the two tissues. The respiration-related parameters in the brain mitochondria were dependent on multiple lipid/phospholipid components, and the process of regulation was complex compared to the liver mitochondrial functions.

Cadmium exposure-induced alterations in the lipid/phospholipids composition of rat brain microsomes and mitochondria

Effects of treatment with a single intra-peritoneal (i.p.) injection (0.84mgCd/kg body weight) of cadmium (Cd) on lipid/phospholipids profiles of rat brain microsomes and mitochondria were examined. At the end of one-week following treatment with Cd the microsomal total phospholipids (TPL) content was unchanged but the cholesterol (CHL) content increased. In one-month Cd-treated group both TPL and CHL contents increased. In one-week Cd-treated group the content of phosphatidylinositol (PI) decreased whereas that of lysophospholipid and phosphatidic acid (PA) increased. In one-month Cd-treated group the sphinghomyelin (SPM) and phosphatidycholine (PC) components increased whereas phosphatidyethanolamine (PE) and PA decreased. In mitochondria, CHL content increased in both Cd-treated groups. The content of TPL decreased only in one-month Cd-treated group. In one-week Cd-treated group the lysophospholipids increased whereas PI and phosphatidylserine (PS) decreased. In one-month group the lysophospholipids, PI and diphosphatidylglycerol (DPG) components decreased whereas PS and PE component increased. The results suggest that exposure to a single dose of Cd has differential and long-lasting effects on lipid/phospholipids profiles of rat brain microsomes and mitochondria.

Effect of treatment with cadmium on kinetic properties of Na+, K+-ATPase and glucose-6-phosphatase activity in rat liver microsomes: A correlative study on influence of lipid/phospholipid make-up

Studies on Cd hepatotoxicity have focused mainly on induction of cytochrome P(450) system and related enzymes. In the present study young adult male rats given a single intra-peritoneal injection of Cd (0.84mg Cd/kg body weight) and effects on kinetic parameters rat liver microsomal Na(+), K(+)-ATPase and G6Pase were evaluated at the end of 1 month and 1 week. The substrate and temperature kinetics parameters were examined and attempts were made to seek correlation with changes in lipid/phospholipid profiles. The Na(+), K(+) ATPase activity decreased only in 1 week Cd-treated group but recovered at the end of 1 month. The activity resolved in two distinct kinetic components in control as well as the experimental groups. In 1 week Cd-treated group the K(m) value of both the components was unchanged, whereas V(max) value decreased. In 1-month Cd-treated group V(max) value only of component I increased. The catalytic efficiency of both the components was not affected in the experimental groups. In 1-week Cd-treated group the energy of activations at high-temperature range (E(H)) and low-temperature range (E(L)) decreased, whereas for 1-month Cd-treated group the energies of activations did not change. The G6Pase activity measured at 37 degrees C was high only in 1-month Cd-treated group. The activity resolved in two kinetically distinguishable components in control as well as in the experimental groups. K(m) value of component I decreased in both the Cd-treated groups. In 1-month Cd-treated group the V(max) value of component II increased. The catalytic efficiency of G6Pase was unchanged despite changes in K(m) and V(max). In 1-week Cd-treated group the E(H) and E(L) decreased, whereas only E(L) showed decrease in 1-month Cd-treated group. Cholesterol (CHL) content increased in both the Cd-treated groups. Content of lysophospholipid (Lyso), spinghomyelin (SPM) and phosphatidic acid (PA) increased, whereas phosphatidylcholine (PC) and phosphatidylserine (PS) decreased in 1-week Cd-treated group. In 1-month Cd group the Lyso, SPM, and PC increased while PC, phosphatidylethanolamine (PE) and PA decreased. In conclusion, Cd has short-term effects on microsomal Na(+), K(+)-ATPase which are reversed by the end of 1 month and that G6Pase does not seem to be a target of Cd insult.

Dehydroepiandrosterone Treatment Alters Lipid/Phospholipid Profiles of Rat Brain and Liver Mitochondria

Stimulation of mitochondrial function following treatment with dehydroepiandrosterone (DHEA) has been demonstrated. Since the activity of several electron transport chain components is dependent on specific lipid/phospholipid components, we examined the effects of DHEA treatment (0.1-2.0 mg/kg body weight for 7 consecutive days) on lipid/phospholipids profiles of rat brain and liver mitochondria. In the brain mitochondria, contents of both total phospholipids (TPL) and cholesterol (CHL) increased. The major effect on phospholipids profile was increase in the contents of lysophospholipids (Lyso) and sphingomyelin (SPM) component followed by phosphatidylinositol (PI) and phosphatidylserine (PS). The contents of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and diphosphatidylglycerol (DPG) were not affected. At the higher dose (2.0 mg) the observed effects declined. The TPL and CHL contents of liver mitochondria were generally unchanged by DHEA treatment. Under this condition the content of PI and PS increased. The contents of other phospholipid components were not changed. Our results suggest that the observed changes may complement the function of electron transport chain components.

Thyroid Hormone-Induced Alterations in Membrane Structure-Function Relationships: Studies on Kinetic Properties of Rat Kidney Microsomal Na+,K+-ATPase and Lipid/Phospholipid Profiles

The effects of thyroidectomy (Tx) and subsequent treatment with 3,5,3′-triiodothyronine (T3) or combined replacement therapy (TR) with T3 and thyroxine (T4) on the substrate and temperature kinetics properties of Na+,K+-ATPase and lipid/phospholipid makeup of rat kidney microsomes were examined. Enzyme activity was somewhat high in the hypothyroid (Tx) animals and increased significantly following T3 treatment, while TR treatment caused a decrease. In the Tx and T3 groups enzyme activity resolved in two kinetic components, while in the TR group the enzyme showed allosteric behavior up to 0.5 mm ATP concentration. The Km and Vmax values of both the components decreased in Tx animals without affecting the catalytic efficiency. T3 treatment caused a significant increase in the Vmax of both the components, with a significant increase in the catalytic efficiency, while the Km values were not upregulated. The TR regimen lowered the Km and Vmax of component II but improved the catalytic efficiency. Thyroid status-dependent changes were also noted in the temperature kinetics of the enzyme. Regression analysis revealed that changes in the substrate and temperature kinetics parameters correlated with specific phospholipid components.

Diabetic Modulation of the Temperature Kinetics Properties of Cytochrome Oxidase Activity in Rat Brain Mitochondria

The effects of alloxan-diabetes and subsequent treatment with insulin on temperature kinetics properties of cytochrome oxidase activity from rat brain mitochondria were examined. The enzyme activity decreased only at the late stage of diabetes which was not normalized by insulin treatment; however at early stage of diabetes hyper-stimulation occurred. In the control animals the Arrhenius plot was chair shaped with three energies of (E1, E2 and E3) and two phase transition temperatures (Tt1 and Tt2). At early diabetic stage the Arrhenius plot became biphasic and E1 and E2 decreased; insulin treatment reversed chair-shaped pattern with increase in E2. These changes correlated with transient changes in the phospholipids profiles especially decreased acidic phospholipids. The temperature kinetics parameters were minimally affected at the late stage of diabetes or by insulin treatment. Thus at the late stage the brain tissue seems to have readjusted to its insulin homeostasis.

Thyroid Hormone Treatments Differentially Affect the Temperature Kinetics Properties of FoF1 ATPase and Succinate Oxidase as well as the Lipid/Phospholipid Profiles of Rat Kidney Mitochondria: A Correlative Study

AbstractEffect of thyroidectomy (Tx) and subsequent treatment with 3,5,3′-triiodo-l-thyronine (T3) or replacement therapy (TR) with T3 + l-thyroxine (T4) on the temperature kinetics properties of FoF1 adenosine triphosphatase (ATPase, ATP synthase, H+-translocating ATP synthase EC 3.6.3.14) and succinate oxidase (SO) and on the lipid/phospholipid makeup of rat kidney mitochondria were examined. Tx lowered ATPase activity, which T3 treatment restored. SO activity was unchanged in Tx but decreased further by T3 treatment. TR restored both activities. The energies of ATPase activation in the high and low temperature ranges (EH and EL) increased in the Tx and T3 animals with decrease in phase transition temperature (Tt). TR restored EH and EL but not Tt to euthyroid levels. EH and EL of SO decreased in Tx animals. T3 and TR restored EH whereas EL was restored only in the TR group; Tt increased in both groups. Total phospholipid and cholesterol contents decreased significantly in Tx and T3-treated animals. In Tx animals, sphingomyelin (SPM) and phosphatidylcholine (PC) components decreased, while phosphatidylserine (PS) and diphosphatidylglycerol components increased. T3 and TR treatments caused decreases in SPM, phosphatidylinositol and PS. PC and phosphatidylethanolamine (PE) increased in the T3 group. TR resulted in increased lysophospolipids and PE. Changes in kinetic parameters of the two enzymes were differently correlated with specific phospholipid components. Both T3 and TR regimens were unable to restore normal membrane structure-function relationships.

Thyroidal regulation of substrate kinetics properties of cytochrome oxidase in rat liver mitochondria

Effects of thyroidectomy (Tx) and subsequent treatment with 3,5,3’-triiodothyronine (T3), and combined treatment (TR) with T3 + thyroxine (T4) on substrate kinetics properties of cytochrome oxidase of rat liver mitochondria were examined. Tx resulted in lowering of cytochromes content with decrease in the enzyme activity, and Km and Vmax. T3 and TR regimens restored the cytochromes contents and the Vmax values to normal. In control, T3 and TR groups the enzyme activity resolved in two kinetic components; in Tx group three kinetic components were evident. The Km values for all components decreased significantly in the experimental groups with concomitant increase in catalytic efficiency, Kcat/Km. Significant alterations in the contents of total phospholipid and of cholesterol were noted while the changes in the phospholipids composition were only of restricted nature. Regression analysis revealed that total phospholipid, cholesterol and phosphatidylcholine, phosphatidylethanolamine play significant role in fine tuning the enzyme activity.