Functions of cytochrome c in regulation of electron transfer and protein folding.

Cytochrome c, a "mobile electron carrier" of the mitochondrial respiratory chain, also occurs in detectable amounts in the cytosol, and can receive electrons from cytochromes present in endoplasmic reticulum and plasma membranes as well as from superoxide and ascorbate. The pigment was found to dissociate from mitochondrial membranes in liver and kidney when rats were subjected to heat exposure and starvation, respectively. Treating cytochrome c with hydroxylamine gives a partially deaminated product with altered redox properties; decreased stimulation of respiration by deficient mitochondria, increased reduction by superoxide, and complete loss of reducibility by plasma membranes. Mitochondria isolated from brown adipose tissue of cold-exposed rats are found to be sub-saturated with cytochrome c. The ability of cytochrome c to reactivate reduced ribonuclease is now reinterpreted as a molecular chaperone role for the hemoprotein.

Polyvanadate acts at the level of plasma membranes through aadrenergic receptor and affects cellular calcium distribution and some oxidation activities.

The activities of calcium-stimulated respiration, calcium uptake, a-glycerophosphate dehydrogenase and rates of oxidation in state 3 and of H2O2 generation, were found to increase and that of pyruvate dehydrogenase decrease in mitochondria isolated from livers of rats administered intraperitoneally or perfused with polyvanadate. Phenoxybenzamine, an antagonist of a−adrenergic receptor, effectively prevented these changes. It was also found that perfusion of the liver with polyvanadate reproduced one of the best characterized events of a-adrenergic activation-stimulation of protein kinase C in plasma membrane accompanied by its decrease in cytosol. These experiments indicate for the first time the α-adrenergic mimetic action of polyvanadate.

Changes in thyroid hormone and insulin status of the brown adipose tissue of cold acclimated rats on short term exposure to heat.

Acclimation of rats to cold caused 45% increase in the concentration of triidothyronine (T3) and 35% increase in the concentration of thyroxine (T4) in serum. Exposure of cold-acclimated rats to heat (12 hr, 37 degrees C) failed to decrease the concentrations of thyroid hormones in circulation. The concentration of T3 in brown adipose tissue (BAT) increased almost 10-fold on cold acclimation. Iodothyronine deiodinase activity also registered 3-fold increase. Exposure of cold-acclimated animals to heat caused decrease in the concentration of T3 in BAT without appreciably affecting T4 concentration. In liver tissue, the changes in hormone concentrations were quite small compared to those in BAT. On thyroidectomy or when fed with propyl thiouracil, rats could not survive exposure to the cold. The concentration of insulin in circulation showed small increase, while that in the tissues showed significant decrease on acclimation of rats to the cold. The concentration of the hormone in BAT registered significant increase on exposure of cold-acclimated animals to heat (12 hr, 37 degrees C). The increase in liver was marginal. The temperature-dependent response of T3 indicates an important role for this hormone in rapid physiological response in BAT.

Increase in alpha-glycerophosphate dehydrogenase and other oxidoreductase activities of hepatic mitochondria on administration of vanadate to the rat.

Liver mitochondria isolated from vanadate-administered rats showed increased (20-25%) rates of oxidation of both NAD(+)-linked substrates and succinate. Respiratory control index and ADP/O were unaffected by the treatment. Dormant and uncoupler-stimulated ATPase activity also was not affected by vanadate administration. Membrane-bound, electron-transport-linked dehydrogenase activities (both NAD(+)- and succinate-dependent) increased by 15-20% on vanadate treatment. Mitochondrial alpha-glycerophosphate dehydrogenase activity increased by 50% on vanadate administration. The above effects of vanadate on oxidoreductase activities could be prevented by the prior administration of antagonists to alpha-adrenergic receptors. Substrate-dependent H2O2 generation by mitochondria also showed an increase on vanadate administration.

Increase in hepatic ubiquinone on administration of diethylhexyl phthalate to the rat.

It is shown for the first time that the content of ubiquinone of liver increases (2·5 fold) on dietary administration of the widely-used industrial plasticizer diethylhexyl phthalate to the rat. The increase is localized almost entirely in mitochondria in which the concentration of the quinone per mg protein is 1·7 times the control. Incorporation of the radioactive precursor (acetate) reveals that the biosynthesis of ubiquinone is increased in the livers of plasticizer-administered animals. The rate of degradation is not altered.