Metabolism and incorporation into glycerolipids of exogenous 18∶3(n−3) and 18∶3(n−6) by MDCK cells

The extent to which exogenous 18∶3(n−3) and 18∶3(n−6) were desaturated and elongated and the degree to which they and their derivatives altered the unsaturation index of cell glycerolipids were compared using clone 4 MDCK cells grown in lipid- and serum-free medium. Despite differences in the degree of unsaturation of the individual polyunsaturated fatty acids produced from 18∶3(n−3) or 18∶3(n−6), the unsaturation index of phospholipids increased similarly from 0.7 in control cells grown in serum- and lipid-free medium to ca. 1.6 in those supplemented with fatty acid. The added fatty acids had little effect on cell growth. The conversion of 18∶3(n−6) to 20∶3(n−6) and 20∶4(n−6) was more rapid than that of 18∶3(n−3) to 20∶4(n−3) and 20∶5(n−3). No significant quantities of 20∶3(n−3) or 18∶4(n−3) were noted. When both 18∶3 isomers were supplied simultaneously, marked differences in the amounts of some species of n−3 and n−6 polyunsaturated fatty acids were observed. The presence of 18∶3(n−6) and/or its derivatives suppressed levels of 20∶4(n−3) and 20∶5(n−3), perhaps through inhibition of the Δ6 and Δ5 desaturases responsible for their synthesis from 18∶3(n−3). Similarly 18∶3(n−3), and/or its longer more unsaturated derivatives, diminished the formation of 20∶4(n−6) from 18∶3(n−6). No marked effect on the products derived from elongation alone were observed.     

Occurrence of 22∶3n−9 and 22∶4n−9 in the lipids of the topminnow (Poeciliopsis lucida) hepatic tumor cell line,PLHC-1

The lipids of the hepatic tumor cell line, PLHC-1, from the topminnow (Poeciliopsis lucida), were found to contain considerable amounts of a range of n−9 polyunsaturated fatty acids despite culture in serum containing significant amounts of essential fatty acids. The structural identity of all the n−9 polyunsaturated fatty acids was confirmed by gas chromatography/mass spectrometry. Of particular interest, PLHC-1 cell total lipid contained 1.9% of 22∶3n−9 and 3.3% of 22∶4n−9. As the culture medium contained virtually no n−9 polyunsaturated fatty acids, these fatty acids are all formed by the PLHC-1 cells, presumably form 18∶1n−9. The 22∶3n−9 and 22∶4n−9 are presumably formed by processes of elongation and “Δ4” desaturation of Mead acid, 20∶3n−9, present at over 11% in fatty acids of total lipid. Both 22∶3n−9 and 22∶4n−9 were primarily located in phosphatidylserine (4.1 and 8.5% respectively) and, to a lesser extent, in phosphatidylethanolamine (2.2 and 6.5%, respectively), in common with the C₂₂ derivatives of the n−3 and n−6 series, whereas 20∶3n−9 was preferentially located in phosphatidylinositol (31.2%). The results establish that long-chain polyunsaturated fatty acids of the n−9 series can be formed in vertebrate tissue other than in conditions of classical essential fatty acid deficiency.     

Functional and ultrastructural effects of essential fatty acid deficiency in kidney epithelial cells

Madin-Darby canine kidney (MDCK) epithelial cells were grown in culture medium supplemented with 1% fetal bovine serum (FBS) to provide a cell culture model of essential fatty acid deficiency (EFAD). 5,8,11-Eicosatrienoic acid (20∶3n−9) accumulated in cellular phospholipids, and arachidonic acid (20∶4) decreased. A large increase in cellular cholesterol/phospholipid ratio was observed. Hemicyst formation was greatly reduced from normal levels in the EFAD-MDCK cells. Scanning and transmission electron microscopy revealed that EFAD-MDCK were much flatter than their normal counterparts. They had much less dense surface microvilli, mitochondria and other organelles were very sparse, except in the perinuclear area, and much of the peripheral cytoplasm was amorphous. The EFAD was rapidly reversed by the addition of as little as 10 μM linoleic or arachidonic acid to the medium. Cells supplemented with 10% FBS, the usual culture condition, displayed borderline EFAD, with intermediate levels of 20∶3n−9 and 20∶4 and hemicyst formation. These studies suggest that EFAD reduces water and electrolyte transport in renal tubular epithelium.     

Polyenoic acid metabolism in cultured human skin fibroblasts

The incorporation of [1-¹⁴C]linoleic acid, and [1-¹⁴C]linoleic acid into cellular lipids of cultured human skin fibroblasts was studied. Cultured cells took up both labeled fatty acids at nearly the same rate and incorporated them into a variety of lipid classes. At the end of 1 hr incubation with [1-¹⁴C]linoleic acid, radioactivity was found in the triacylglycerol (TG) and choline phosphoglyceride (CPG) pools preferentially. Incorporation into the TG fraction decreased rapidly, while the uptake into CPG, serine phosphoglyceride (SPG), and ethanolamine phosphoglyceride (EPG) fractions increased progressively with longer incubation times. Similar results were obtained with [1-¹⁴C]linoleic acid as precursor. At the end of 24 hr, desaturation and chain elongation of 18∶3 n−3 was more extensive than conversion of 18∶2 n−6 to higher polyenoic acids. During pulse-chase experiments with either fatty acid precursor, the incorporated radioactivity was progressively lost from cellular lipids, particularly from the TG and CPG fractions, but continued to increase in the SPG and EPG pools. The similar labeling pattern of cellular phospholipids with linoleic or linolenic acids, and data from pulse-chase studies suggest that a direct transfer of fatty acids from CPG to EPG is a likely pathway in fibroblast cultures. Incorporation into the EPG pool during the pulse-chase experiments paralleled extensive desaturation and elongation of linoleic acid into 20∶4 n−6, and 22∶4 n−6; and of linolenic acid into 22∶5 n−3 and 22∶6 n−3.     

Effect of dietary n−3 and n−6 polyunsaturated fatty acids on lipid-metabolizing enzymes in obese rat liver

This study was designed to examine whether n−3 and n−6 polyunsaturated fatty acids at a very low dietary level (about 0.2%) would alter liver activities in respect to fatty acid oxidation. Obese Zucker rats were used because of their low level of fatty acid oxidation, which would make increases easier to detect. Zucker rats were fed diets containing different oil mixtures (5%, w/w) with the same ratio of n−6/n−3 fatty acids supplied either as fish oil or arachidonic acid concentrate. Decreased hepatic triacylglycerol levels were observed only with the diet containing fish oil. In mitochondrial outer membranes, which support carnitine palmitoyltransferase I activity, cholesterol content was similar for all diets, while the percentage of 22∶6n−3 and 20∶4n−6 in phospholipids was enhanced about by 6 and 3% with the diets containing fish oil and arachidonic acid, respectively. With the fish oil diet, the only difference found in activities related to fatty acid oxidation was the lower sensitivity of carnitine palmitoyltransferase I to malonyl-CoA inhibition. With the diet containing arachidonic acid, peroxisomal fatty acid oxidation and carnitine palmitoyltransferase I activity were markedly depressed. Compared with the control diet, the diets enriched in fish oil and in arachidonic acid gave rise to a higher specific activity of aryl-ester hydrolase in microsomal fractions. We suggest that slight changes in composition of n−3 or n−6 polyunsaturated fatty acids in mitochondrial outer membranes may alter carnitine palmitoyltransferase I activity.     

Influence of environmental medium on fatty acid composition of human cells: Leukocytes and fibroblasts

Fibroblasts in culture and leukocytes have been widely used to study fatty acid and lipoprotein cellular metabolism. The present investigations were designed to study the role of nutritional and environmental factors on lipid metabolism in these two types of cells. Leukocytes freshly isolated from human blood and fibroblasts cultured in media enriched in human serum (HS) have relatively similar fatty acid distributions. However, more important differences are observed in fibroblasts cultured in media enriched with HS or with fetal bovine serum (FBS). It is obvious that the quantity and quality of fatty acids are very different in FBS and HS, but intracellular regulation ensures relative homogeneity of saturated (SFA) and monounsaturated fatty acids (MUFA) in the cells, particularly in phospholipids. The first modifications induced by different media (FBS or HS) are detected on cellular growth; the differences seem to be due more to the fatty acid (FA) quantitative supply than to the FA quality of each culture medium. The major modifications in FA composition induced by different culture media concern the polyunsaturated fatty acids (PUFA) of phospholipids, especially the n−6 family. The intracellular linoleic acid level depends on the level in the medium, but intracellular n−6 metabolite levels depend both on the level in the medium and on the growth state of the cells. The n−3 family seems to be less affected by the quality of the medium in our experiment, and the cells maintain a stable docosahexaenoic acid (22∶6n−3) level. A higher content of the n−3 family in the medium induces a higher level of eicosa-or docosapentaenoic acid, rather than docosahexaenoic acid itself. Finally, the FA quality of the medium influences the cellular PUFA content but, with a low FA quantitative supply, the FA quality of the medium has less influence on the cellular PUFA quality, and apparently has no effect on the SFA content of phospholipids. Modification of the quantitative supply of the medium and of the quality of the cells (strain and growing state) are more important for the distribution of SFA and MUFA in the neutral lipids of the cells.     

Incorporation of n−3 fatty acids into WB-F344 cell phospholipids inhibits gap junctional intercellular communication

In this investigation, we demonstrate that rat liver epithelial (WB-F344) cells grown in medium supplemented with n−3 fatty acids (FA) results in the inhibition of gap junctional intercellular communication (GJIC). Cells incubated for 48 hr in medium containing 50 μM α-linolenate (18∶3n−3) resulted in a 60% inhibition of GJIC, compared to control cells, while treatment with γ-linolenate (18∶3n−6) had no effect. Supplementation with octadecatetraenoate (18∶4n−3), eicosapentaenoate (20∶5n−3), and docosahexaenoate (22∶6n−3), inhibited GJIC by 42%, 28%, and 18%, respectively. Incubation with each of the n−3 FA markedly increased the total n−3 FA content of cellular phospholipids (PL). Growing cells in medium containing 50 μM arachidonate (20∶4n−6) plus 50 μM 18∶3n−3 partially attenuated the inhibition of GJIC induced by 18∶3n−3. The mechanism by which n−3 FA inhibit GJIC remains to be determined.     

Specific modifications of phosphatidylinositol and nonesterified fatty acid fractions in cultured porcine cardiomyocytes supplemented with n-3 polyunsaturated fatty acids

Mechanisms for the antiarrhythmic effect of n−3 polyunsaturated fatty acids (PUFA) are currently being investigated using isolated cardiac myocytes. It is still not known whether the incorporation of n−3 PUFA into membrane phospholipids is a prerequisite for its protective action or if n−3 PUFA exert antiarrhythmic effects in their nonesterified form as demonstrated by recent studies. Adult porcine cardiomyocytes were grown in media supplemented with arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). After 24 h, analysis of total lipids showed that the myocytes were enriched with the respective fatty acids compared to control cells. Large proportions of all three fatty acids supplemented (69% AA, 72% DHA, and 66% EPA) remained unesterified. Fatty acid analysis of total phospholipids (PL) revealed that the incorporation of EPA and DHA, though small, was significantly different (P<0.05) from that of the control cells. The PL fraction was further separated into phosphatidylinositol (Pl), phosphatidylethanolamine, phosphatidylcholine, and phosphatidylserine to study the pattern of incorporation of the fatty acids in these fractions. It became apparent that EPA and DHA were selectively incorporated into the Pl fraction. This study demonstrates that in adult porcine cardiomyocytes, the n−3 PUFA supplementation selectively modulates two important lipid fractions, nonesterified fatty acid and Pl, which were implicated in the mechanisms of prevention of cardiac arrhythmias.     

Quantitative effects of dietary polyunsaturated fats on the composition of fatty acids in rat tissues

A method combining data on fatty acid composition into subsets is used to illustrate general relative competitive selectivities in the metabolic and transport events that maintain fatty acid compositions in tissue lipids and to minimize differences among tissues or species in the amount of individual fatty acids. Fatty acid compositions of triglycerides and phospholipids in several tissues of the rat were maintained with simple relationships between the exogenous n−3 and n−6 dietary polyunsaturated fatty acids and the endogenous n−7 and n−9 types of fatty acid. The general pattern of fatty acids in triglycerides was similar for liver, plasma and adipose tissue, averaging about 30% as saturated acids, 67% as 16- and 18-carbon unsaturated acids and only about 2% as 20- and 22-carbon highly unsaturated acids. The tissues maintained a linear relationship between the amount of 18-carbon polyunsaturated fatty acids in the diet and in the tissue triglycerides, with the proportionality constant for 18∶3n−3 being 60% of that for 18∶2n−6. The total phospholipids of liver, plasma and red blood cells maintained about 45% of the fatty acids in the form of saturated fatty acids and 20–30% as 20- and 22-carbon highly unsaturated fatty acids irrespective of very different proportions of n−3, n−6 and n−9 types of fatty acids. In all three tissues, the 20-carbon highly unsaturated fatty acids of the n−3, n−6 and n−9 type were maintained in a competitive hyperbolic relationship with apparent EC₅₀ values for dietary 18∶2n−6 and 18∶3n−3 near 0.1% of dietary calories. The consistent quantitative relationships described in this study illustrate an underlying principle of competition among fatty acids for a limited number of esterification sites. This approach may be useful in predicting the influence of diet upon tissue levels of the substrates and antagonists of eicosanoid biosynthesis.     

Lipids of cultured hepatoma cells: II. Effect of media lipids on cellular phospholipids

Minimal deviation hepatoma cells were cultured in a modified Swim's 77 medium supplemented with decreasing amounts of serum, lipid-free serum, and lipid-free serum containing added palmitic or linoleic acids. Cellular phospholipids were extracted and the class distribution determined quantitatively. The fatty acid composition of each phospholipid class was determined, and the percentages from cells grown on each of the various media were compared. Cellular phospholipid class and fatty acid compositions differed from media compositions, indicating that intact serum phospholipids are not incorporated into cellular structures. Phosphatidylcholine percentages decreased as the media serum and lipid levels decreased, while phosphatidylinositol and phosphatidylethanolamine percentages increased. Sphingomyelin of cells grown in medium containing added linoleic acids contained a high level of a 24∶2 acid. All classes, except sphingomyelin, contained elevated levels of 18∶1 acid and decreased levels of polyunsaturated fatty acids, relative to normal rat liver. Cells cultured on lipid-free medium did not contain increased concentrations of 20∶3 acid, suggesting that this hepatoma cell cannot desaturate monoenoic acids. Phosphoglycerides of cells, grown on lipid-free medium, had the highest monoene fatty acid concentration, whereas those cells grown on media containing added linoleic acid had the lowest concentrations, suggesting that linoleate may inhibit or regulate monoenoic acid biosynthesis in this cell. These mass data also demonstrate that monoenoic fatty acid biosynthesis in this cultured hepatoma cell responds to dietary changes.     

Chain elongation of polyunsaturated fatty acids by vascular endothelial cells: Studies with arachidonate analogues

This study has utilized radiolabeled analogues of arachidonic acid to study the substrate specificity of elongation of long-chain polyunsaturated fatty acids. Human umbilical vein endothelial cells were incubated for 2–72 hr in medium supplemented with 0.9–2.6 μM [¹⁴C]fatty acid, and cellular glycerolipids were analyzed by gas-liquid chromatography with radioactivity detection. Elongation of naturally occurring C₂₀ polyunsaturated fatty acids occurred with eicosapentaenoate (20∶5(n−3))>Mead acid (20∶3(n−9))>arachidonate (20∶4(n−6)). Chain length markedly influenced the extent of elongation of 5,8,11,14-tetraenoates (18∶4>19∶4>20∶4>21∶4); effects of initial double bond position were also observed (6,9,12,15–20∶4>4,7,10,13–20∶4. Neither 5,8,14- nor 5,11,14–20∶3 was elongated to the extent of 5,8,11–20∶3. Differences between polyunsaturated fatty acids were observed both in the initial rates and in the maximal percentages of elongation, suggesting that the content of cellular C₂₀ and C₂₂ fatty acids may represent a balance between chain elongation and retroconversion. Umbilical vein endothelial cells do not exhibit significant desaturation of either 22∶4(n−6) or 22∶5(n−3). By contrast, incubation with 5,8,11,14-[¹⁴C]18∶4(n−4) resulted in formation of both [¹⁴C]20∶5(n−4) and [¹⁴C]22∶5(n−4). The respective time courses for the appearances of [¹⁴C]22∶5(n−4) and [¹⁴C]20∶5(n−5) suggests Δ6 desaturation of [¹⁴C]22∶4(n−4) rather than Δ4 desaturation of [¹⁴C]20∶4(n−4).     

Modulation of cyclic nucleotide phosphodiesterase by dietary fats in rat heart

Feeding oils of different fatty acid composition modifies the fatty acid composition of cardiac membrane phospholipids, thereby inducing changes in cardiac contractility and altering response of adenylate cyclase to catecholamines. In the present study, the effect of such dietary manipulations on cyclic nucleotide phosphodiesterase, which is involved in the control of cyclic nucleotide intracellular levels and in the control of cardiac contractility, was investigated. Rats were fed either a saturated fatty acid-enriched diet (8 weight percent [%] coconut oil +2% sunflower oil), an n−6 fatty acid-enriched diet (10% sunflower oil) or an n−3 fatty acid-enriched diet (8% fish oil +2% sunflower oil). The fatty acid composition of cardiac phospholipids, as well as the nonesterified fatty acid content of heart were markedly altered by the diets. The 18∶2n−6 and 20∶4n−6 content of cardiac phospholipids was markedly (−49%) depressed by fish oil as compared with sunflower oil feeding, but the nonesterified fatty acid level of heart membrane was lowest in coconut oil-fed rats. In addition, fish oil feeding more drastically depressed the n−6/n−3 fatty acid ratio in the nonesterified fatty acid pool than in cardiac phospholipids. Cyclic AMP phosphodiesterase activity was the lowest in both the particulate and soluble fractions of heart from rats fed sunflower oil, whereas cyclic GMP phosphodiesterase activity was not altered by the diets. Cyclic AMP phosphodiesterase activity was decreased by 18 and 12% in heart membranes of the sunflower oil group as compared to that of the coconut oil and fish oil groups, respectively. In heart cytosol, the activity decreased by 30% when compared with the activity of the coconut oil group. Additionalin vitro experiments showed that polyunsaturated fatty acids were more potent inhibitors of cyclic AMP phosphodiesterase than saturated fatty acids. These results suggest that polyunsaturated fatty acid-enriched diets might decrease heart cyclic AMP phosphodiesterase activity by increasing non-esterified polyunsaturated fatty acids, especially those of the n−6 series, but more complex and indirect mechanisms are very likely to be involved.     

Interaction of (n−3) and (n−6) fatty acids in desaturation and chain elongation of essential fatty acids in cultured glioma cells

Recent research in various biological systems has revived interest in interactions between the (n−6) and (n−3) essential fatty acids. We have utilized cultured glioma cells to show that linolenic acid, 18∶3(n−3), is rapidly desaturated and chain elongated; 20∶5(n−3) is the major product and accumulates almost exclusively in phospholipids. We examined effects of various (n−6), (n−3), (n−9) and (n−7) fatty acids at 40 μM concentration on desaturation and chain elongation processes using [1-¹⁴C]18∶3(n−3) as substrate. In general, monoenoic fatty acids were without effect. The (n−6) fatty acids (18∶2, 18∶3, 20∶3, 20∶4 and 22∶4) had little effect on total product formed. There was a shift of labeled product to triacylglycerol, and in phospholipids, slightly enhanced conversion of 20∶5 to 22∶5 was evident. In contrast, 22∶6(n−3) was inhibitory, whereas 20∶3(n−3) and 20∶5(n−3) had much less effect. At concentrations <75 μM, all acids were inhibitory. Most products were esterified to phosphatidylcholine, but phosphatidylethanolamine also contained a major portion of 20∶5 and 22∶5. We provide a condensed overview of how the (n−6) and (n−3) fatty acids interact to modify relative rates of desaturation and chain elongation, depending on the essential fatty acid precursor. Thus, the balance between these dietary acids can markedly influence enzymes providing crucial membrane components and substrates for biologically active oxygenated derivatives.     

Effect of low levels of dietary fish oil on fatty acid desaturation and tissue fatty acids in obese and lean rats

The effect of very low levels of dietary long-chain n−3 fatty acids on Δ6 desaturation of linoleic acid (18∶2n−6) and α-linolenic acid (18∶3n−3), and on Δ5 desaturation of dihomo-γ-linolenic acid (20∶3n−6), in liver microsomes and its influence on tissue fatty acids were examined in obese and lean Zucker rats and in Wistar rats. Animals fed for 12 wk a balanced diet containing ca. 200 mg of long-chain polyunsaturated n−3 fatty acids per 100 g of diet were compared to those fed the same amount of α-linoleic acid. Low amounts of long-chain n−3 fatty acids greatly inhibited Δ6 desaturation of 18∶2n−6 and Δ5 desaturation of 20∶3n−6, while Δ6 desaturation of 18∶3n−3 was not inhibited in Zucker rats and was even stimulated in Wistar rats. Inhibition of the biosynthesis of long-chain n−6 fatty acids was reflected in a decrease in arachidonic acid (20∶4n−6) content of serum lipids when fasting, and also in the phospholipid fatty acids of liver microsomes. On the contrary, heart and kidney phospholipids did not develop any decrease in 20∶4n−6 during fish oil ingestion. Docosahexaenoic acid (22∶6n−3), present in the dietary fish oil, was increased in serum lipids and in liver microsome, heart, and kidney phospholipids.     

Accumulation of (n−9)-eicosatrienoic and docosatrienoic acids in human fibroblast phospholipids

An essential fatty acid (EFA) deficiency-like profile of fatty acids has been observed in HF-1 human skin fibro-blasts cultured at clonal densities in MCDB 110 and 0.4% fetal bovine serum (FBS). The profile was characterized by an accumulation of 16∶1n−7, 18∶1n−9, 20∶3n−9 and 22∶3n−9, a reduction of n−6 fatty acids and a reduction in total polyunsaturated fatty acids (PUFA). The fatty acid composition of sphingomyelin (SPH), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylinositol (PI) and phosphatidylethanolamine (PE) was determined and, except for SPH, each displayed an EFA deficiency-like profile. The triene to tetraene ratio (20∶3n−9/20∶4n−6) ranged from 5.3 in PI to 0.9 in PE. In addition, the highest percentage of 20∶3n−9 was present in the PI and the highest percentage of 22∶3n−9, in PE. Other human fibroblasts (normal, transformed and at different population doubling number levels [PDL] were grown under the same conditions and were found to display triene to tetraene ratios (20∶3n−9/20∶4n−6) in total cellular lipids ranging from 0.7 to 4.5. The accumulation of 20∶3n−9 and 22∶3n−9 is due primarily to the existence of a basal nutrient medium (MCDB 110) that allows for the rapid clonal growth of human fibroblasts at reduced serum levels (0.4%). This culture procedure can be exploited to further elucidate various aspects of lipid metabolism in human fibroblasts. Fatty acids are abbreviated as number of carbon atoms: number of double bonds, followed by an n-number to designate the position of the first double bond with respect to the methyl carbon. Thus, Mead acid is 20∶3n−9 and its elongation product is 22∶3n−9.     

Trans-monoenoic and polyunsaturated fatty acids in phospholipids of aVibrio species of bacterium in relation to growth conditions

AVibrio species of bacterium known to contain the polyunsaturated fatty acid 20∶5n−3 was grown in both freshwater and seawater media at 5 and 20°C and examined for adaptive changes in lipid composition. Phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), together with a smaller proportion of nonesterified fatty acids (NEFA), comprised almost all the lipid under all growth conditions examined. Temperature had a more pronounced effect than the salinity of the medium on lipid composition. The proportion of PE in total lipid was always higher at 5 than at 20°C. Conversely, the proportion of NEFA was lower at 5 than 20°C whereas that of PG was not altered. The levels of saturated fatty acids in total lipid, PE and PG were all decreased by growth at 5°C. No differences were observed with respect to growth temperature in the levels ofcis 16∶1n−7, the principal monoenoic fatty acid in both PE and PG.Trans 16∶1n−7 was found to comprise 12.8–15.2% of fatty acids in PE and PG of bacteria grown at 5°C but only 4.4–8.5% of phospholipid fatty acids in bacteria cultured at 20°C. Regardless of medium composition, a reduction in growth temperature from 20 to 5°C also caused the proportions of 20∶5n−3 to increase from around 0.8 to 4.4% in PE and from around 4 to 20% in PG. The simultaneous occurrence oftrans 16∶1n−7 and 20∶5n−3 is unique to thisVibrio species of bacterium. The increased proportions of both these fatty acids with decreasing temperature suggest that they have a role in retailoring biomembrane phospholipids during temperature acclimation of the bacterium.     

Linoleic acid-induced fatty acid changes in platelet and aorta of the rat: Effect of age and cholesterol

The influence of age and cholesterol on polyunsaturated fatty acids (PUFa) levels was studied in young and old male Sprague-Dawley rats. Animals were fed a fat-free diet supplemented with 10% (by wt) safflower oil with or without 1% cholesterol for 8 wk. As a result of cholesterol feeding, proportions of linoleic acid (18∶2n−6) and dihomo-γ-linolenic acid (30∶3n−6) were increased and and that of arachidonic acid (20∶4n−6) was decreased in the liver and platelet phospholipids in 64-wk-old rats, suggesting inhibitory effects of cholesterol on 20∶4n−6 synthesis from 18∶2n−6. The prominent age-dependent effect on the levels of PUFA was a retention of C−22 n−3 PUFA, accompanied by decreased C−22 n−6 PUFA and increased 20∶3n−6 in the liver and platelet phospholipids. Ratio of 20∶3n−6/20∶4n−6 increased in 64-wk-old rats regardless of dietary cholesterol, suggesting depressed Δ5-desaturase with age. In aorta phospholipids, 20∶3n−6 content and 20∶3n−6/20∶4n−6 ratio increased with cholesterol supplementation, but not with age. These results suggest that changes of PUFA composition of platelet phospholipids with age are closely linked with changes in liver phospholipids. The 20∶4n−6 content in both platelet and aorta phospholipids is kept constant, despite other n−6 and n−3 PUFA being affected by age.     

Effects of dietary n−3 polyunsaturated fatty acids on phospholipid composition and calcium transport in mouse cardiac sarcoplasmic reticulum

The effects of dietary n−3 and n−6 polyunsaturated fatty acids on the fatty acid composition of phospholipid, Ca⁺⁺· Mg⁺⁺ ATPase and Ca⁺⁺ transport activities of mouse sarcoplasmic reticulum were investigated. Mice were fed a 2 weight percent fat diet containing either 0.5 weight percent ethyl esters of 18∶3n−3, 20∶5n−3 or 22∶6n−3 as a source of n−3 polyusaturated fatty acid or 0.5 weight percent safflower oil as a cource of n−6 polyunsaturated fatty acid for 10 days. Olive oil (2 weight percent) was used as a control diet. Although feeding n−6 polyunsaturated fatty acid induced very little modifications of the phospholipid sarcoplasmic reticulum fatty acid composition, feeding n−3 polyunsaturated fatty acid altered it markedly. Inclusion of 18∶−3, 20∶5n−3 or 22∶6n−3 in the diet caused an accumulation of 22∶6n−3, which replaced 20∶4n−6 and 18∶2n−6 in phospholipid sarcoplasmic reticulum. The saturated fatty acids were significantly increased with a concurrent reduction of 18∶1n−9. These changes in the fatty acid composition resulted in a decrease in the values of the n−6/n−3 polyunsaturated fatty acid ratio and a decrease in the ratio of 20 carbon to 22 carbon fatty acids esterified in the phospholipid sarcoplasmic reticulum. This was associated with a decrease in Ca⁺⁺ uptake by n−3 polyunsaturated fatty acid enriched sarcoplasmic reticulum vesicles as compared with n−6 fatty acid and control diet sarcoplasmic reticulum vesicles. However, neither the affinity for Ca⁺⁺ nor the maximal velocity of ATP hydrolysis activity of Ca⁺⁺·MG⁺⁺ ATPase were altered by the different diets. The data suggest that the incorporation of 22∶6n−3 and/or the decrease of 20∶4n−6 plus 18∶2n−6 in the phospholipid sarcoplasmic reticulum may affect the membrane lipid bilayer structure and make it more permeable to Ca⁺⁺.     

Fatty acid composition of late embryonic and early postnatal rat brain

The fatty acid (FA) composition and distribution in a variety of phospholipids (PL) and neutral lipids (NL) at two discrete stages during the embryonic rat brain development were investigated. Over 96% of the FA were acylated into fetal brain PL at embryonic day 17 after the peak of neuronal proliferation and at embryonic day 20, one day prior to delivery. Phosphatidylcholine constituted approximately 60% of the total PL pool, phosphatidylethanolamine (PE) 30%, phosphatidylserine (PS) 6%, and phosphatidylinositol (PI) 4%. The diacylglycerols and triacylglycerols constituted 1–2% of the fetal brain lipids. α-Linolenic acid (18∶3n−3) and linoleic acid (18∶2n−6) were found in very low amounts in all fetal brain PL and NL. The percentage of the n−6 polyunsaturated FA, consisting of arachidonic acid (AA), 22∶4n−6 and 22∶5n−6, remained unchanged in all the fractions, except in Pl, in which the proportion of AA increased. The concentration of docosahexaenoic acid (DHA) increased with age in all the fractions, with the bulk of accumulation accounted for by its increase in PE and, to a lesser extent, in PS. This finding suggests a “DHA accretion spurt” during the last three days of pregnancy.     

Fatty acid composition of the adipose tissue and yolk lipids of a bird with a marine-based diet, the emperor penguin (Aptenodytes forsteri)

The emperor penguin (Aptenodytes forsteri) is an Antarctic seabird feeding mainly on fish and therefore has a high dietary intake of n-3 polyunsaturated fatty acids. The yolk is accumulated in the developing oocyte while the females are fasting, and a large proportion of the fatty acid components of the yolk lipids are derived by mobilization from the female's adipose tissue. The fatty acid composition of the total lipid of the yolk was characterized by high levels of n-3 polyunsaturated fatty acids. However, it differed in several respects from that of the maternal adipose tissue. For example, the proportions of 14∶0, 16∶1n−7, 20∶1n−9, 22∶1n−9, 20∶5n−3, and 22∶6n−3 were significantly greater in adipose tissue than in yolk. Thus adipose tissue lipids contained 7.6±0.3% and 8.0±0.3% (wt% of total fatty acids; mean ±SE; n=5) of 20∶5n−3 and 22∶6n−3, respectively, whereas the yolk total lipid contained 1.6±0.1 and 5.5±0.3% of these respective fatty acids. The proportions of 16∶0, 18∶0, 18∶1n−9, 18∶2n−6, and 20∶4n−6 were significantly lower in the adipose tissue than in the yolk lipids. The proportions of triacylglycerol, phospholipid, free cholesterol, and cholesteryl ester in the yolk lipid were, respectively, 67.0±0.2, 25.4±0.3, 5.3±0.2, and 1.8±0.2% (wt% of total yolk lipid). The proportions of 20∶4n−6, 20∶5n−3, 22∶5n−3, and 22∶6n−3 were, respectively, 5.7±0.3, 2.8±0.2, 1.4±0.1, and 11.7±0.5% in phospholipid and 0.4±0.0, 1.2±0.1, 0.8±0.1 and 3.6±0.3% in triacylglycerol. About 95% of the total vitamin E in the yolks was in the form of α-tocopherol with γ-tocopherol forming the remainder. Two species of carotenoids, one identified as lutein, were present.