Metabolism of (3-(14)C)tryptophan and (2-(14)C)alanine in cattle tissues

In the experiments involving incubation of the liver, brain cortex, muscle and adipose tissues homogenates with [3-14C] tryptophan for an hour 43.2-89.3% of the label was found in proteins, 7.2-47.2%--in lipids, 2.6-9.4%--in CO2. Following incubation of the above-mentioned tissue homogenates with [2-14C] alanine, proteins, lipids and CO2 contain 28.8-49.3%; 22.6-31.9% and 21.6-49.3% of radioactive label, respectively. Radioactivity of lipids synthesized by the homogenates of the investigated tissues from [3-14C] tryptophan and [2-14C] alanine is 23.5-63.5 and 21.1-56.0%, respectively, the radioactivity of CO2 being 1.4-5.1 and 9.3-11.8% of the above-mentioned compounds synthesized from [1-14C] acetate. The results obtained testify to the considerable contribution of [3-14C] tryptophan and [2-14C] alanine to protein synthesis as well as to their involvement in the substrate supply of lipogenesis and energetic processes in various organs and tissues of cattle.     

Reassessment of 14CO2 compartmentation and of [14C]formate oxidation in rat liver

Our previous report (Marsolais, C., Huot, S., David, F., Garneau, M., and Brunengraber, H. (1987) J. Biol. Chem. 262, 2604-2607) had concluded that a fraction of [14C]formate oxidation in liver occurs in the mitochondrion. This conclusion was based on the labeling patterns of urea and acetoacetate labeled via 14CO2 generated from [14C]formate and other [14C]substrates. We reassessed our interpretation in experiments conducted in (i) perifused mitochondria and (ii) isolated livers perfused with buffer containing [14C]formate, [14C]gluconolactone, 14CO2, or NaH13CO3, in the absence and presence of acetazolamide, an inhibitor of carbonic anhydrase. Our data show that the cytosolic pools of bicarbonate and CO2 are not in isotopic equilibrium when 14CO2 is generated in the cytosol or is supplied as NaH14CO3. We retract our earlier suggestion of a mitochondrial site of [14C]formate oxidation.     

The effect of ionizing irradiation on the oxidation of palmitate-1-14C by thymus and liver

New Zealand rabbits, fasted for 12 hours, were subjected to 500 rads of whole-body irradiation. K+-palmitate-1-14C oxidation was assayed with the 600 X g supernatant of thymus and liver homogenates, in the presence of ATP, at various time intervals from irradiation. For a period of 24 hours following irradiation, oxidation by liver preparations was not significantly affected. The rate of oxidation by thymus was decreased to less than one-third of the control value within 12 hours from irradiation and, at 24 hours, was almost completely abolished. Increased ATP concentration could increase only to a small extent the oxidation by thymus preparations of irradiated animals. Oxidation by isolated thymus mitochondria was also inhibited by irradiation. Counting of the water-soluble oxidation products of palmitate-1-14C suggests that the inhibition is not due to the impairment of the reactions of the citric acid cycle. The non-esterified fatty acid concentration of thymus was not altered at 12 hours following irradiation. Esterification of K+-palmitate-l-14C into the thymus lipids was not affected 12 hours after irradiation.     

The L-DOPA-sparing effect of R-(-)-1-(benzofuran-2-yl)-2-propylaminopentane hydrochloride [(-)-BPAP] in reserpine-pretreated rats

R-(-)-1-(Benzofuran-2-yl)-2-propylaminopentane hydrochloride [(-)-BPAP], a highly potent enhancer of impulse propagation-mediated release of catecholamines and serotonin in the brain, and significantly increased the locomotor activity of normal rats at the doses of 0.3 and 1 mg/kg s.c. (P < 0.05), while L-DOPA (200 and 400 mg/kg i.p.) had no significant effect. The locomotor activity of rats simultaneously administered L-DOPA and (-)-BPAP was significantly higher than with (-)-BPAP alone (P < 0.05). In rats pretreated with reserpine (1 mg/kg i.v.), the hypolocomotion was significantly reversed by 400 mg/kg i.p. L-DOPA, or 1 or 3 mg/kg s.c. (-)-BPAP (P < 0.05). Furthermore, the combined administration of subthreshold doses of 200 mg/kg i.p. L-DOPA and 0.3 mg/kg s.c. (-)-BPAP highly potentiated the locomotor activity in the reserpine-pretreated rats. However, (-)-BPAP failed to reverse the hypolocomotion in rats pretreated with reserpine + alpha-methyl-DL-p-tyrosine. Thus, (-)-BPAP was demonstrated to possess the L-DOPA-sparing effect in normal and reserpine-pretreated rats.     

Respiration of 14CO2 by intact animals of various species given l-[1-14C]fucose or d-[1-14C]arabinose

The production of ¹⁴CO₂ from l-[1-¹⁴C]fucose and d-[1-¹⁴C]arabinose has been studied in five mammalian species.Cats, guinea pigs, mice, and rabbits respired about 22% of the label of l[1-¹⁴C]fucose or of d-[1-¹⁴C]arabinose within 6 h after intraperitoneal injection of the sugar. Rats respired only 1.5% of the l-fucose label and 5% of the d-arabinose label in the same time period.Liver homogenates from cat, guinea pig, and rabbit produced significantly more ¹⁴CO₂ from l-[1-¹⁴C]fucose or d-[1-¹⁴C]arabinose than mouse or rat liver homogenates. Unlike those of the other species, guinea pig liver homogenates had very low l-fucose dehydrogenase activity.The results suggest that substantial catabolism of l-fucose and d-arabinose occurs in the tissues of some animal species. Investigators wishing to employ l-fucose as a tracer of glycoprotein metabolism must, therefore, ensure that the species that they employ does not metabolize l-fucose to products interfering with their studies.     

In vitro oxidation of [U-14C] palmitate, [1-14C] and [6-14C] glucose in newborn and adult rats and pigs

Studies have been made on the intensity of oxidation of [U-14C]-palmitate, [1-14C]- and [6-14C]-glucose by slices of the liver and skeletal muscles of new-born, 1-day, 5-day and adult Wistar rats and domestic pigs. It was found that the level of 14CO2 production from these substrates is higher in tissues of rats than in those of pigs. At early stages of ontogenesis, in tissues of both species intensive oxidation of glucose is observed together with oxidation of fatty acids. In the course of ontogenetic development, the intensity of glucose utilization significantly decreases, whereas the level of fatty acid catabolism remains relatively unaffected.     

Effect of dietary alpha-linolenic acid on the conversion of linoleic and gamma-linolenic acids (1-14C) into arachidonates in rats in vivo]

The effects of alpha-linolenic acid (9-12-15 octadecadienoic) upon the conversion in vivo of [1-14C] linoleic acid and of [1-14C] gamma-linolenic acid into arachidonate have been studied in adult rats. The two tracers have been administered by stomach tubing and the amounts of [14C]-radioactivity incorporated into arachidonate in the liver, kidneys and whole rat have been measured 48 h later. Three experiments have been carried out on rats fed on alpha-linolenic acid containing diets prior to the radioactive tubing. In these diets, alpha-linolenic acid was brought either as ethyl ester or in the form of Primor oil (erucic acid free rapeseed oil). In all of them, the ratio alpha-linolenic acid: linoleic acid did not exceed 0.45. Control animals were fed, in the same conditions, ethyl oleate or peanut oil respectively. Comparing the alpha-linolenic acid fed-rats to the control animals, we were able to observe the following results: (1) The exogenous supplies of alpha-linolenic acid used in the diets have not brought about any significant alteration in the amounts (weights) of arachidonic acid present in the liver, kidneys and whole animal. (2) Using [1-14C] linoleic acid as a precursor, the amounts of [14C]-radioactivity incorporated into arachidonate in the same organs as well as in the whole rat have been significantly lowered by dietary alpha-linolenate. (3) alpha-Linolenate, on the contrary, had no significant effect upon the amounts of radioactivity incorporated into hepatic, renal and whole body arachidonate following the administration of [1-14C] gamma-linolenic acid. These results lead to the conclusion that alpha-linolenic acid, when present in the diet of rats at a limited, phyisological level, partly inhibits the desaturation of linoleic acid in vivo but does not affect the subsequent reactions in the biosynthesis of arachidonic acid.     

Conversion of (U-14C)-glycerol, (2-3H)-glycerol and (1-14C)-palmitate into circulating lipoproteins in the rat.

The in vivo formation of labelled very low density lipoproteins (VLDL) from (U-14C)-glycerol, (2-3H)-glycerol and (1-14C)-palmitate was studied in fed female rats. The rate of disappearance of radioactivity from plasma after the i.v. injection with these tracers was similar for (U-14C)-glycerol and (1-14C)-palmitate. With (2-3H)-glycerol, plasma radioactivity at 10 min was lower than with the other substrates although it did not change thereafter. A certain proportion of radioactivity administered as glycerol appeared in plasma lipids, mainly in the VLDL glyceride glycerol fraction, although when (U-14C)-glycerol was the substrate, a considerable portion also appeared in the esterified fatty acids of these lipoproteins. When using (1-14C)-palmitate, practically all the circulating labelled esterified fatty acids appeared in the VLDL fraction, while the labelled free fatty acids appeared in lipoprotein of higher density, presumable free fatty acid-albumin complexes. This data is discussed in terms of the role of the liver in the rapid, continuous cycling of these substrates to yield VLDL-glycerides for their extrahepatic utilization.