The mitochondrial tricarboxylate carrier (TCC) plays an important role in lipogenesis being TCC-responsible for the efflux from the mitochondria to the cytosol of acetylCoA, the primer for fatty acid synthesis. In this study, we investigated the effects of two high-fat diets with different fatty acid composition on the hepatic TCC activity. Rats were fed for 3 weeks on a basal diet supplemented with 15% of either coconut oil (CO), abundant in medium-chain saturated fatty acids, or fish oil (FO), rich in n-3 polyunsaturated fatty acids. Mitochondrial fatty acid composition was differently influenced by the dietary treatments, while no appreciable change in phospholipid composition and cholesterol level was observed. Compared with CO, the TCC activity was markedly decreased in liver mitochondria from FO-fed rats; kinetic analysis of the carrier revealed a decrease of the V max , with no change of the K m . No difference in the Arrhenius plot between the two groups was observed. Interestingly, the carrier protein level and the corresponding mRNA abundance decreased following FO treatment. These data indicate that FO administration markedly decreased the TCC activity as compared with CO. This effect is most likely due to a reduced gene expression of the carrier protein.

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Short-term effects of 3,5-l-diiodothyronine (T2) on lipid biosynthesis were studied in cultured hepatocytes from hypothyroid rats. A comparison with the effects of T3 was routinely carried out. After T2 addition to cell cultures, a distinct stimulation of fatty acid and cholesterol syntheses, measured as incorporation of [1-14C]acetate into these lipid fractions, was observed. The T2 dose-dependent effect on both metabolic pathways, already detectable at 10(-8)-10(-9) M, reached a 2-fold stimulation at 10(-5) M T2. At this concentration, the stimulatory effect was evident within 1 h of T2 addition to the hepatocytes and increased with time up to the length of the experimental period of 4 h. T2 stimulation of lipogenesis was also confirmed by incubating hepatocytes with [3H]H2O, used as an independent index of lipogenic activity. The effects of T2 are rather specific as 3,3',5,5'-tetraiodo-D-thyronine and 3,5-diiodo-L-tyrosine were practically ineffective on both fatty acid and cholesterol synthesis. Analysis of various lipid fractions showed that T2 addition to the cells produced a significant stimulation of the incorporation of newly synthesized fatty acids into both neutral and polar lipids. By comparing the effects induced by T2 with those seen in the presence of T3, it appeared that T2 was able to mimic T3 effects. Experiments conducted in the presence of cycloheximide, a protein synthesis inhibitor, indicated that the T2 stimulatory effect on fatty acid and cholesterol synthesis was essentially independent of protein synthesis.

Cycloheximide drastically reduced the rate of root pressure exudation in detopped tobacco (Nicotiana tabacum L.), and the effect was more pronounced for nitrate salts in the external solution than for some other salts or when the roots were in water. Diurnal periodicity was greatly decreased, and its phase was changed. Effects began within an hour. Cation uptake was reduced by relatively low levels of cycloheximide. The effects of cycloheximide seemed to be reversible. Mild phosphorus deficiency resulted in decreased exudation rates from detopped tobacco and in no response to nitrate. Periodicity of exudation was not greatly affected by phosphorus deficiency, however. Gamma ray irradiation with a 60Co source at levels (up to 40-50 kilo-roentgens) which are considered disruptive of moderately large molecules had relatively little effect on the exudation rate. Higher levels of irradiation, which disrupt most protein molecules, decreased exudation and obscured peri-odicity. The results indicate either that new protein (or peptide) synthesis is needed for the rapid nitrate transport or that the deficiency and inhibitor disrupt cellular membranes. Phosphorus deficiency increased the sensitivity of the plants to inhibition by irradiation of the exudation process.