Precursor For Fatty Acid Synthesis Research Articles

Fatty Acid Oxidation and Insulin Action

Type 2 diabetes is a disease of metabolic dysregulation involving impaired uptake and utilization of glucose, altered lipid metabolism, accumulation of various lipid species in the circulation and in tissues, and disruption of metabolic signaling pathways that regulate insulin secretion from pancreatic islet β-cells. Normal fuel homeostasis involves reciprocal regulation of glucose and lipid catabolism. Fundamental contributions to our understanding of the interplay between these two key groups of metabolic fuels came from the work of Randle (1), who demonstrated that increased rates of fatty acid oxidation in the fasted state lead to suppression of glucose oxidation and activation of gluconeogenesis, thereby preserving blood glucose for use by the brain and central nervous system. Conversely, the transition from the fasted to the fed state involves a coordinated shift from fatty acid to glucose oxidation. A key element in the latter switch, as elegantly demonstrated by the work of McGarry (2), is the glucose-induced rise in malonyl CoA, which inhibits fatty acid oxidation via direct binding to and allosteric inhibition of carnitine palmitoyltransferase-1 (CPT-1), the rate limiting enzyme for transport of cytosolic long-chain acyl CoA molecules into the mitochondria for oxidation. The multifaceted roles of malonyl CoA as a key glucose-derived metabolite, an allosteric inhibitor of fatty acid oxidation, and a biosynthetic precursor for fatty acid synthesis has led to a series of recent studies investigating the effects of manipulating this metabolite in various tissues. A bonus of such experiments is the opportunity to assess the physiological impact of enhanced or diminished fat oxidation in different cell types and in whole animals. In this issue of Diabetes …

C/EBPα is required for lung maturation at birth

Epithelial cells lining the peripheral lung synthesize pulmonary surfactant that reduces surface tension at the air-liquid interface. Lack of surfactant lipids and proteins in the lungs causes respiratory distress syndrome, a common cause of morbidity and mortality in preterm infants. We show that C/EBPalpha plays a crucial role in the maturation of the respiratory epithelium in late gestation, being required for the production of surfactant lipids and proteins necessary for lung function. Deletion of the Cebpa gene in respiratory epithelial cells in fetal mice caused respiratory failure at birth. Structural and biochemical maturation of the lung was delayed. Normal synthesis of surfactant lipids and proteins, including SP-A, SP-B, SP-C, SP-D, ABCA3 (a lamellar body associated protein) and FAS (precursor of fatty acid synthesis) were dependent upon expression of the C/EBPalpha in respiratory epithelial cells. Deletion of the Cebpa gene caused increased expression of Tgfb2, a growth factor that inhibits lung epithelial cell proliferation and differentiation. Normal expression of C/EBPalpha required Titf1 and Foxa2, transcription factors that also play an important role in perinatal lung differentiation. C/EBPalpha participates in a transcriptional network that is required for the regulation of genes mediating perinatal lung maturation and surfactant homeostasis that is necessary for adaptation to air breathing at birth.

Comparative Aspects of Lipid Metabolism: Impact on Contemporary Research and Use of Animal Models

The emerging obesity crisis and consequent concerns for corrective measures and appropriate public policy have stimulated research into causes, prevention, remediation, and health consequences of obesity and associated maladies. Such research areas include eating behavior, appetite control, and food intake regulation as well as the regulation of lipid metabolism, cardiovascular function, endocrine function, and dyslipidemia states utilizing various animal models and cell culture systems. Although the liver has a central role in lipid/fatty acid synthesis and glucose is the precursor for de novo fatty acid synthesis in rodents and humans, in many other species, adipose tissues are the primary sites of lipogenesis. In addition, many species utilize acetic acid as a precursor for fatty acid synthesis. This fundamental difference in the site of fatty acid synthesis and the pattern of consequent lipid trafficking influences overall animal lipid metabolism and the role of regulatory hormones and transcription factors. Researchers utilizing various animal species in targeted biomedical research should be aware of these species differences when interpreting their data. In addition, many animal species are used for food production, recreational, and companion purposes. Understanding the lipid metabolism regulatory mechanisms of such species from a comparative perspective is important for the proper nutrition and health of these animals.

Lactate carbon does not enter the sugars of lipopolysaccharide when gonococci are grown in a medium containing glucose and lactate: implications in vivo

In media containing glucose, lactate stimulates the metabolism of gonococci at concentrations that simulate conditions in vivo. Nuclear magnetic resonance (NMR) spectroscopy of 13C-labelled lipids obtained from gonococci grown in a synthetic medium with 13C-labelled lactate and unlabelled glucose (culture A), 13C-labelled glucose alone (culture B) or 13C-labelled glucose and unlabelled lactate (culture C) showed lactate carbon was not present in glycerol/ethanolamine residues of lipids from culture A. This indicated that, in the presence of glucose, lactate gluconeogenesis is shut down. Hence, the stimulation of metabolism could result from the production of extra energy because lactate is used solely for conversion to acetyl-CoA, the precursor of fatty acid synthesis and the components of the tricarboxylic acid cycle. In this paper, additional evidence for lack of gluconeogenesis has been sought using a different approach. The carbohydrate moieties of lipopolysaccharide (LPS) have been examined for lactate carbon after gonococci were grown with lactate and glucose. Two methods were used: NMR spectroscopy of 13C-labelled lipopolysaccharide purified from the three cultures described above showed that, in the presence of glucose, lactate carbon, in contrast to glucose carbon, was not in the carbohydrate moiety. Also, 14C-labelled lactate was added to a culture containing unlabelled glucose and lactate (culture A) and [ 14C]glucose to cultures containing unlabelled glucose without unlabelled lactate (culture B) and with unlabelled lactate (culture C). When LPS samples purified from these cultures were subjected to hydrazinolysis, the ratio of the radioactivity of water-soluble products (carbohydrate moieties) to those of chloroform-soluble products (fatty acids) was much lower when [ 14C]lactate was used in culture A, than when [ 14C]glucose was used in cultures B and C. Thus, in the presence of glucose, lactate carbon, unlike glucose carbon, is incorporated predominantly into fatty acids of LPS, not into its carbohydrate moieties. There is no doubt, therefore, that gluconeogenesis is shut off when lactate is present with glucose and there is a consequent stimulation of metabolism. This probably occurs in vivo on mucous surfaces, where gonococci are surrounded by a mixture of glucose and lactate in the secretions.

Understanding in vivo carbon precursor supply for fatty acid synthesis in leaf tissue.

The principal supply of carbon precursors for fatty acid synthesis in leaf tissue has been a much debated topic, with some experiments suggesting a direct supply from the C3 products of photosynthetic carbon fixation and colleagues suggesting the utilization of free acetate (for which concentrations in leaves in the range of 0.05-1.4 mM have been reported). To address this issue we first reassessed the in vivo rate of fatty acid synthesis using a new method, that of [13C]carbon dioxide labeling of intact Arabidopsis plants with the subsequent analysis of fatty acids by gas chromatography-mass spectrometry (GC-MS). This method gave an average value of 2.3 mmoles carbon atoms h-1 mg chlorophyll-1 for photosynthetic tissues. The method was extended by isotopic dilution analysis to measure the rate of fatty acid synthesis in the dark. There was negligible fatty acid synthesis (< 5% of the rate in the light) in the dark. In addition, the method allowed an estimate of the absolute rate of fatty acid degradation of about 4% of the total fatty acid content per day. With the in vivo rate of fatty acid synthesis in the light defined, if the bulk tissue acetate concentration available for fatty acid synthesis is 1 mM, this acetate pool can sustain fatty acid synthesis for approximately 60 min. When the leaves of Arabidopsis, barley and pea were given a 5 min pulse of [14C]carbon dioxide, the label rapidly appeared in fatty acids with a lag phase of less than 2-3 min. Continuous labeling with [14C]carbon dioxide, for up to 1 h, showed a similar result. Furthermore, 14C-label in free acetate was less than 5% of that in fatty acids. In conclusion, these data suggest that either the bulk pool of acetate is not involved in fatty acid synthesis or the concentration of acetate must be less than 0.05 mM under strong illumination.

Why do mitochondria synthesize fatty acids? Evidence for involvement in lipoic acid production.

The function of acyl carrier protein (ACP) in mitochondria isolated from pea leaves has been investigated. When pea leaf mitochondria were labeled with [2-14C] malonic acid in vitro, radioactivity was incorporated into fatty acids, and, simultaneously, ACP was acylated. [1-14C]Acetate was much less effective as a precursor for fatty acid synthesis, suggesting that mitochondria do not possess acetyl-CoA carboxylase. The incorporation of radioactivity from [2-14C]malonate into fatty acids and the labeling of ACP were inhibited by cerulenin and required ATP and Mg2+. These findings indicate that plant mitochondria contain not only ACP, but all enzymes required for de novo fatty acid synthesis. Over 30% of the radioactive products from pea mitochondria labeled with [2-14C]malonate were recovered in H protein, which is a subunit of glycine decarboxylase and contains lipoic acid as an essential constituent. In similar experiments, the H protein of Neurospora mitochondria was also labeled by [2-14C]malonate. The labeling of pea H protein was inhibited by addition of cerulenin into the assay medium. Together, these findings indicate that ACP is involved in the de novo synthesis of fatty acids in plant mitochondria and that a major function of this pathway is production of lipoic acid precursors.

Initial steps of sophoroselipid biosynthesis by Candida bombicola ATCC 22214 grown on glucose.

Candida bombicola ATCC 22214 produces the glycolipid sophoroselipid when cultivated on a medium with glucose as the sole carbon source. Under phosphate-limiting conditions the product yield rises from 0.033 to 0.143 and the specific product formation rate rises from 0.004 h-1 to 0.007 h-1. Enhanced sophoroselipid synthesis is initiated by the decline of the specific activities of NAD- and NADP-dependent isocitrate dehydrogenase (EC 1.1.1.41 and 1.1.1.42) to 2% and 0% of the initial activities respectively. Constantly high specific activity of citrate synthase (EC 4.1.3.7) causes an accumulation of isocitrate and citrate in the mitochondria. Both acids are transported into the cytosol where citrate is cleaved by ATP: citrate lyase (EC 4.1.3.8) giving rise to acetyl-CoA, the precursor of fatty acid synthesis. The ATP: citrate lyase is unaffected by different energy charges; the apparent K(m) values for coenzyme A, ATP and citrate are 23 microM, 250 microM and 256 microM respectively. NADPH for fatty acid synthesis might be generated by further metabolism of oxaloacetate, the other product of the citrate-cleaving reaction, by oxidation of the isocitrate by the cytosolic NADP-dependent isocitrate dehydrogenase or via the hexose monophosphate shunt. A possible explanation for sophoroselipid formation during exponential growth is given.

Inhibition of fatty acid synthesis in Escherichia coli in the absence of phospholipid synthesis and release of inhibition by thioesterase action.

The effects of inhibition of Escherichia coli phospholipid synthesis on the accumulation of intermediates of the fatty acid synthetic pathway have been previously investigated with conflicting results. We report construction of an E. coli strain that allows valid [14C]acetate labeling of fatty acids under these conditions. In this strain, acetate is a specific precursor of fatty acid synthesis and the intracellular acetate pools are not altered by blockage of phospholipid synthesis. By use of this strain, we show that significant pools of fatty acid synthetic intermediates and free fatty acids accumulate during inhibition of phospholipid synthesis and that the rate of synthesis of these intermediates is 10 to 20% of the rate at which fatty acids are synthesized during normal growth. Free fatty acids of abnormal chain length (e.g., cis-13-eicosenoic acid) were found to accumulate in glycerol-starved cultures. Analysis of extracts of [35S]methionine-labeled cells showed that glycerol starvation resulted in the accumulation of several long-chain acyl-acyl carrier protein (ACP) species, with the major species being ACP acylated with cis-13-eicosenoic acid. Upon the restoration of phospholipid biosynthesis, the abnormally long-chain acyl-ACPs decreased, consistent with transfer of the acyl groups to phospholipid. The introduction of multicopy plasmids that greatly overproduced either E. coli thioesterase I or E. coli thioesterase II fully relieved the inhibition of fatty acid synthesis seen upon glycerol starvation, whereas overexpression of ACP had no effect. Thioesterase I overproduction also resulted in disappearance of the long-chain acyl-ACP species. The release of inhibition by thiosterase overproduction, together with the correlation between the inhibition of fatty acid synthesis and the presence of abnormally long-chain acyl-ACPs, suggests with that these acyl-ACP species may act as feedback inhibitors of a key fatty acid synthetic enzyme(s).

Physiological modulation of the uptake and fate of glucose in brown adipose tissue.

Glucose utilization indices (GUI values) and rates of fatty acid synthesis in interscapular brown adipose tissue (IBAT) varied during the diurnal cycle in virgin and late-pregnant rats permitted unrestricted access to food. In virgin rats, peak GUI values and lipogenic rates were observed at the end of the dark (feeding) phase, but were not sustained during the light phase. Whereas peak GUI values were comparable with those observed during re-feeding after 24 h starvation, maximum rates of IBAT fatty acid synthesis in virgin rats during the diurnal cycle were only approx. 25% of those measured during re-feeding after 24 h starvation. Despite hyperphagia, GUI values during the diurnal cycle in late-pregnant rats fed ad libitum were generally lower than those of age-matched virgin controls. The percentage of pyruvate dehydrogenase complex present in the active form (PDHa) was also significantly decreased. Suppression of GUI and PDHa was not parallelled by suppression of fatty acid synthesis. IBAT GUI values in late-pregnant rats during chow re-feeding ad libitum after 24 h starvation were only 25% of those of corresponding virgin controls, and stimulation of fatty acid synthesis was also dramatically attenuated. The suppression of IBAT GUI values after re-feeding in pregnancy was not due to depletion of GLUT 4 protein. The results are discussed in relation to the importance of glucose as a precursor for fatty acid synthesis in IBAT.

Is Acetylcarnitine a Substrate for Fatty Acid Synthesis in Plants?

Long-chain fatty acid synthesis from [1-14C]acetylcarnitine by chloroplasts isolated from spinach (Spinacia oleracea), pea (Pisum sativum), amaranthus (Amaranthus lividus), or maize (Zea mays) occurred at less than 2% of the rate of fatty acid synthesis from [1-14C]acetate irrespective of the maturity of the leaves or whether the plastids were purified using sucrose or Percoll medium. [1-14C]-Acetylcarnitine was not significantly utilized by highly active chloroplasts rapidly prepared from pea and spinach using methods not involving density gradient centrifugation. [1-14C]Acetylcarnitine was recovered quantitatively from chloroplast incubations following 10 min in the light. Unlabeled acetyl-L-carnitine (0.4 mM) did not compete with [1-14C]acetate (0.2 mM) as a substrate for fatty acid synthesis by any of the more than 70 chloroplast preparations tested in this study. Carnitine acetyltransferase activity was not detected in any chloroplast preparation and was present in whole leaf homogenates at about 0.1% of the level of acetyl-coenzyme A synthetase activity. When supplied to detached pea shoots and detached spinach, amaranthus, and maize leaves via the transpiration stream, 1 to 4% of the [1-14C]acetylcarnitine and 47 to 57% of the [1-14C]acetate taken up was incorporated into lipids. Most (78-82%) of the [1-14C]acetylcarnitine taken up was recovered intact. It is concluded that acetylcarnitine is not a major precursor for fatty acid synthesis in plants.

Lipogenesis in rat brown adipocytes. Effects of insulin and noradrenaline, contributions from glucose and lactate as precursors and comparisons with white adipocytes.

1. Brown adipocytes were isolated from the interscapular depot of male rats maintained at approx. 21 degrees C. In some experiments parallel studies were made with white adipocytes from the epididymal depot. 2. Insulin increased and noradrenaline decreased [U-14C]glucose incorporation into fatty acids by brown adipocytes. Brown adipocytes differed from white adipocytes in that exogenous fatty acid (palmitate) substantially decreased fatty acid synthesis from glucose. Both noradrenaline and insulin increased lactate + pyruvate formation by brown adipocytes. Brown adipocytes converted a greater proportion of metabolized glucose into lactate + pyruvate and a smaller proportion into fatty acids than did white adipocytes. 3. In brown adipocytes, when fatty acid synthesis from [U-14C]glucose was decreased by noradrenaline or palmitate, incorporation of 3H2O into fatty acids was also decreased to an extent which would not support proposals for extensive recycling into fatty acid synthesis of acetyl-CoA derived from fatty acid oxidation. 4. In the absence of glucose, [U-14C]lactate was a poor substrate for lipogenesis in brown adipocytes, but its use was facilitated by glucose. When brown adipocytes were incubated with 1 mM-lactate + 5 mM-glucose, lactate-derived carbon generally provided at least 50% of the precursor for fatty acid synthesis. 5. Both insulin and noradrenaline increased [U-14C]glucose conversion into CO2 by brown adipocytes (incubated in the presence of lactate) and, in combination, stimulation of glucose oxidation by these two agents showed synergism. Rates of 14CO2 formation from glucose by brown adipocytes were relatively small compared with maximum rates of oxygen consumption by these cells, suggesting that glucose is unlikely to be a major substrate for thermogenesis. 6. Brown adipocytes from 6-week-old rats had considerably lower maximum rates of fatty acid synthesis, relative to cell DNA content, than white adipocytes. By contrast, rates of fatty acid synthesis from 3H2O in vivo were similar in the interscapular and epididymal fat depots. Expressed relative to activities of fatty acid synthase or ATP citrate lyase, however, brown adipocytes synthesized fatty acids as effectively as did white adipocytes. It is suggested that the cells most active in fatty acid synthesis in the brown adipose tissue are not recovered fully in the adipocyte fraction during cell isolation. Differences in rates of fatty acid synthesis between brown and white adipocytes were less apparent at 10 weeks of age.

Fatty acid and acylglycerol glycerol synthesis in isolated liver cells from foetal lambs

1. 1. The rates of fatty acid and acylglycerol glycerol synthesis from acetate, glucose and l-lactate in isolated liver cells from foetal lambs at about 30 and 3 days before birth are reported. The total rates of fatty acid synthesis from endogenous precursors and added substrates, measured using 3H 2O, are also reported for these liver cells. 2. 2. The total rate of fatty acid synthesis did not differ at the two ages. Of the added substrates, acetate and l-lactate were better precursors for fatty acid synthesis than glucose under the conditions used. Endogenous precursors, probably glycogen glucose, made a substantial contribution to fatty acid synthesis. 3. 3. Results are compared with previous studies of fatty acid synthesis in adipose tissue from foetal lambs.

297 EFFECT OF INTRAUTERINE GROWTH RETARDATION (IUGR) ON FATTY ACID SYTHESIS

The effects of IUGR on fetal triacylglycerol fatty acid (FA) synthesis was studied in 26 pregnant rats by 3H2O injection. IUGR was induced by uterine artery ligation on one horn and litters from the opposite horn served as controls. The specific activity of FA at 19 days of gestation in the liver was lower in the IUGR fetus, as compared to controls, 44.6±13 vs. 87.9±10 cpm/μmole FA, (p<.05). At 21 days of gestation, the specific activity of FA remains lower in the IUGR group, (40.6±7.8 vs. 75.8±9.8 cpm/μmole p<.05). In lung, the specific activity of FA was 25.7±5.4 cpm/μmole in IUGR fetuses as compared to 46.6±9 cpm/μmole in controls at 19 days (p<.05). This difference persisted at 21 days (32.2±2.4 vs. 42.6±3.0 cpm/μmole in IUGR and controls, respectively, p<.05).The total fatty acid content per ogran for both liver and lung was also reduced in the IUGR group. However, when expressed per unit organ weight, (gram) the IUGR fetuses have a significantly higher FA content at 21 days than the controls. This observation is probably related to a proportionally greater reduction in the accretion rates of substrates other than fat during growth. The data indicates that fetal growth retardation induced by placental vascular insufficiency is associated with reduced fat synthesis probably on the basis of reduced placental transfer of precursors for fatty acid synthesis. Similar phenomenon may be present in human IUGR infant.

Effect of substrate interactions on in vitro fatty acid synthesis in adipose tissue of normal and genetically obese mice

Fatty acid synthesis was measured in vitro in pieces of adipose tissue from lean and obese-hyperglycaemic (ob/ob) mice, using 14C-glucose or 14C-lactate and 3H2O to obtain absolute rates of total fatty acid synthesis. In the presence of lipoprotein-triglyceride (2.5 mumol/l) metabolic interaction occurred which decreased glucose incorporation into fatty acids by 30% in lean mouse tissue, but not in obese mouse tissue. In the absence of added insulin, the contribution of glucose to total fatty acid synthesis was 69% in obese mouse tissue, significantly lower than the value of 87% obtained in lean mouse tissue. Insulin increased the contribution of glucose to total synthesis in both lean and obese mouse tissues, although the value in obese mouse tissue (83%) remained lower than the value in lean mouse tissue (100%). Lactate was not a major precursor for fatty acid synthesis. When both lactate (2 mmol/l) and glucose (15 mmol/l) were present, the contribution of lactate to total fatty acid synthesis was not increased in obese mouse tissue, suggesting that even in the presence of insulin, about 30% of the carbon was provided by intracellular precursors.

Esterification of exogenous and endogenous fatty acids by rat adipocytes in vitro

Rat adipocytes were used in vivo to compare the esterification of exogenous fatty acids and fatty acids formed de novo from glucose or acetate. Pure single fatty acids added to the medium were esterified at comparable rates but marked differences were observed when the same acids were supplied as components of a fatty acid mixture of a composition similar to that in the tissue. Fatty acids synthesised de novo from acetate by adipocytes in a medium containing high concentrations of acetate were located predominantly in diacylglycerols. The effect was most marked with adipocytes from older rats and was enhanced by the presence of exogenous long-chain fatty acids. Exogenous oleic acid was esterified predominantly into triacylglycerols at all concentrations of acetate. No such accumulation of endogenously-synthesised fatty acids in diacylglycerols occurred when glucose was the precursor for fatty acid synthesis. The diacylglycerols formed were almost entirely of the sn-1,2-configuration.

Effect of Level of Feed Intake on Lactate and Acetate Metabolism and Lipogenesis In Vivo in Sheep

Experiments were conducted to investigate the relative importance of acetate and lactate as precursors for fatty acid synthesis in vivo in sheep fed ad libitum [3.3 Mcal of metabolizable energy (ME)/day] or a restricted level of feed intake (1.7 Mcal of ME/day). Tritiated water (3H2O) was also used to estimate fatty acid synthesis from all carbon precursors rather than from a specific carbon precursor. In experiment 1, the amount of tritium (3H) incorporated into adipose tissue fatty acids increases with time up to 6 hours after intravenous 3H2O injection in perirenal, mesenteric, omental, shoulder, rump, backfat and abdominal adipose tissue from ad libitum fed sheep. The rate of incorporation did not differ significantly between tissue sites. The 3H2O equilibrated rapidly with body water and the plasma 3H2O specific activity did not change significantly for 3 hours after 3H2O injection. Plasma acetate turnover rate was not influenced by ad libitum versus restricted feeding and averaged 2.99 ± 0.28 mmole/hour/kg of body weight. Plasma lactate turnover was decreased significantly in restricted versus ad libitum fed sheep and averaged 0.99 ± 0.01 and 1.76 ± 0.07 mmoles/hour/kg of body weight, respectively. Restricting feed intake for 14 days decreased the total weight of perirenal, mesenteric, and omental adipose tissue masses compared to ad libitum fed sheep. Total 3H incorporated into fatty acids in restricted fed sheep was only 11% to 13% of that in the ad libitum fed sheep in perirenal and omental adipose tissue; treatment differences in 3H incorporation were much smaller in the total carcass adipose tissue. On the basis of the [14C]lipid relative specific activity, lactate was incorporated into carcass adipose tissue fatty acids at 38% of the rate for acetate in ad libitum fed sheep and only 16% and 21% of the rate for acetate in perirenal and omental adipose tissue. Lactate appears to have been more important as a lipid precursor in ad libitum fed than in restricted fed sheep.

Ketone bodies serve as important precursors of brain lipids in the developing rat.

AbstractKetone bodies are readily oxidized for energy by extrahepatic tissues. Since oxidation of ketone bodies produces acetyl coenzyme A (AcCoA), and hence could be an important source of immediate precursors for fatty acid synthesis, we investigated, in whole‐brain homogenates of developing rats, the preferential utilization of [3‐14C]acetoacetate (AcAc), [3‐14C]β‐hydroxybutyrate (β‐OHB), and [U‐14C]glucose for production of CO2 and lipids, including phospholipids, glycerides, cholesterol, and free fatty acids. Throughout the postnatal period, the rate of AcAc oxidation was 2–3 and 2–6 times the rate for β‐OHB and glucose, respectively. The eynthesis of lipids from AcAc was 7‐ to 11‐fold higher than from glucose. The brain's capacity for lipid synthesis from β‐OHB was similar to that from AcAc during the first 8 days of life; however, during the next 10 days, the synthesis of lipids from β‐OHB decreased to 60% of AcAc‐dependent synthesis. The high rate of lipid synthesis from ketone bodies was accompanied by increased activities of cytoplasmic acetoacetyl CoA synthetase and acetoacetyl CoA thiolase in the developing brain. During the entire postnatal development, the proportion of radioactivity claimed by lipids vs. CO2 from [3‐14C]AcAc was 44–62% vs. 38–56%; from [3‐14C]β‐OHB, 50–81% vs. 19–50%; and from [U‐14C]glucose, 14–43% vs. 57–86%. Phospholipids accounted for more than two‐thirds of total lipids synthesized from either ketone bodies or glucose, while diglycerides plus cholesterol and free fatty acids accounted for most of the remainder. Addition of glucose to the incubation medium did not alter lipid production from AcAc throughout the suckling period, but moderately depressed energy production in the brain of 16‐ to 20‐day‐old rats. It is clear that in cell‐free preparations from the brain of developing rats, ketone bodies are preferred over glucose as precursors for both energy and lipids, mainly phospholipids. These results suggest that ketone bodies are important for the growth and development of the brain.

A possible role for malonyl-CoA in the regulation of hepatic fatty acid oxidation and ketogenesis.

Studied on the oxidation of oleic and octanoic acids to ketone bodies were carried out in homogenates and in mitochondrial fractions of livers taken from fed and fasted rats. Malonyl-CoA inhibited ketogenesis from the former but not from the latter substrate. The site of inhibition appeared to be the carnitine acyltransferase I reaction. The effect was specific and easily reversible. Inhibitory concentrations were in the range of values obtained in livers from fed rats by others. It is proposed that malonyl-CoA functions as both precursor for fatty acid synthesis and suppressor of fatty acid oxidation. As such, it might be an important element in the carbohydrate-induced sparing of fatty acid oxidation.

Metabolism of Liver Triacylglycerols in Rats Tube-Fed a Threonine-Devoid Diet

Rats tube-fed a diet devoid of threonine accumulated triacylglycerols in their livers, starting on the third day of the diet. The fatty acid composition of the accumulated lipid and the contribution of novo synthesized fatty acids to the lipid accumulation, as determined with tritiated water as a radioactive precursor for fatty acid synthesis, suggested that an increased hepatic de novo synthesis of fatty acids is not a major factor for the development of this liver lipid accumulation. The metabolism of intravenous injected 3H-oleic acid, the Triton-induced hyperlipemia and the activity of lipoprotein lipase in adipose tissue was also studied. None of these studies revealed any significant difference between the threonine-deficient and control rats. It is concluded that the hepatic triacylglycerol accumulation in the threonine-deficient rats does not result from any gross abnormality in the rate of liver triacylglycerol formation or secretion to the plasma. It is suggested that a possible causative mechanism is a derangement in the metabolism of the storage pool of liver triacylglycerols.

Différences de nature dans les systèmes de transport extra-mitochondrial des précurseurs de la synthèse des acides gras chez le rat et la souris

Summary A comparative study of the extramitochondrial carriers of precursors for fatty acid synthesis in rats and mice has been made. Animals treated with amino-oxyacetate (AOA) received intravenous injections of either [2-3H] acetate or [2-14C] pyruvate and killed 5 min thereafter. In rats, the radioactivity found in liver fatty acids increases when the precursor is [2-3H] acetate but not when it is [2-14C] pyruvate. On the contrary, an inhibition of fatty acid synthesis is observed in mice mainly after administration of [2-14C] pyruvate. Administration of AOA results in inhibition of gluconeogenesis from [2-14C] pyruvate in rats but a stimulation is observed in mice. AOA does not modify the intracellular distribution of neo-synthesized fatty acids in rat liver. The activity of hepatic citrate cleavage enzyme of rats is twice that of mice. The incorporation of tritium of 3H2O (which is not submitted to dilution phenomena) in the fatty acids of rats and mice has also been studied as a function of time. AOA inhibits fatty acid synthesis in mouse adipose tissue and liver at all time intervals studied. However, only a slight inhibition was observed in rat liver. Varying concentrations of pyruvate do not influence the stimulatory effect of AOA on the incorporation of [2-3H] acetate but the administration of citrate suppresses it. The results are discussed and it is concluded that in the liver, citrate is a more efficient carrier of acetyl CoA in rats than in mice and that extramitochondrial carriers of precursors for fatty acid synthesis are different in rats and mice.