Effects of two Avtemia diets with different contents of polyunsaturated fatty acids on the lipid composition of larvae of Atlantic herring (Clupea harengus

Atlantic herring larvae ( Clupea harengus ) were fed two enriched Artemia diets with different contents of (n-3) highly unsaturated fatty acids (HUFA), one containing low levels of 20: 5(n-3) and no 22: 6(n-3), the other containing substantial levels of both 20: 5(n-3) and 22: 6(n-3). After 30 days of culture, fatty acid compositions of lipid classes in the heads, bodies and eyes of the larvae were analysed. Fish fed Artemia with the low (n-3) HUFA diet lacking 22: 6(n-3) had lower amounts of total (n-3)HUFA and, in particular, of 22:6(n-3) in individual phospholipids and total neutral lipids of heads, bodies and eyes as compared to fish fed Artemia with high levels of (n-3)HUFA. The amount of 22: 6(n-3) in the fatty acids of phosphatidyl-ethanolamine of eyes was particularly susceptible to dietary depletion. The implications of these findings are discussed, particularly in relation to dietary requirements for 22: 6(n-3) during development of neural tissue in predatory fish iarvae.     

Conservation of Docosahexaenoic Acid in Rod Outer Segments of Rat Retina During n-3 and n-6 Fatty Acid Deficiency

We investigated the mechanism by which rat retina conserves docosahexaenoic acid during essential fatty acid deficiency. Weanling female albino rats were fed diets containing either 10% by weight hydrogenated coconut oil, safflower oil, or linseed oil for 15 weeks. Plasma and rod outer segment (ROS) membranes were prepared for fatty acid and phospholipid molecular species analysis. In addition, retinas were removed for morphometric analysis. We found the following: (1) Plasma phospholipids and cholesterol esters from coconut oil, safflower oil, and linseed oil diet groups were enriched in 20:3(n-9), 20:4(n-6), and 20:5(n-3), respectively. The levels of these 20-carbon fatty acids in the ROS, however, were only slightly affected by diet. (2) The fatty acids and molecular species of ROS phospholipids from the safflower oil and coconut oil groups showed a selective replacement of 22:6(n-3) with 22:5(n-6), as evidenced by a reduction of the 22:6(n-3)-22:6(n-3) molecular species and an increase in the 22:5(n-6)-22:6(n-3) species. (3) The renewal rate of ROS integral proteins, determined by autoradiography, was 10% per day for each diet group. (4) Morphometric analysis of retinas showed no differences in the outer nuclear layer area or in ROS length between the three groups. We conclude that the conservation of 22:6(n-3) in ROS is not accomplished through reductions in the rate of membrane turnover, the total amount of ROS membranes, or in the number of rod cells. The retina may conserve 22:6(n-3) through recycling within the retina or between the retina and the pigment epithelium, or through the selective uptake of 22-carbon polyunsaturated fatty acids from the circulation.     

Effect of long-term supplementation with arachidonic or docosahexaenoic acids on sperm production in the broiler chicken

The possibility was investigated that dietary supplementation of the male chicken with long-chain polyunsaturated fatty acids of the n-6 and n-3 series may prevent the decrease in sperm output that normally occurs by 60 weeks of age. From 26 weeks of age, birds were raised on wheat-based diets supplemented with either maize oil (rich in linoleic acid, 18:2n-6), arasco oil (rich in arachidonic acid, 20:4n-6) or tuna orbital oil (rich in docosahexaenoic acid, 22:6n-3). The effects of the last two oils were investigated at two levels of vitamin E supplementation (40 and 200 mg kg(-1) feed). By 60 weeks of age, there was a small increase in the proportion of the main polyunsaturate of chicken sperm phospholipid, docosatetraenoic acid 22:4n-6, in chickens fed arasco oil diet compared with chickens given the maize oil diet, an effect that was potentiated at the higher dietary intake of vitamin E. Supplementation with tuna orbital oil significantly reduced the proportions of 20:4n-6 and 22:4n-6 in the sperm phospholipid and increased the proportion of 22:6n-3. The diet supplemented with tuna orbital oil and the lower level of vitamin E markedly depleted vitamin E from the tissues of the birds and decreased the concentration of vitamin E in the semen; these effects were largely prevented by the higher level of vitamin E in the diet. The susceptibility of semen to lipid peroxidation in vitro was increased in chickens fed arasco and tuna orbital oils with 40 mg vitamin E kg(-1) feed, but was reduced when 200 mg vitamin E kg(-1) feed was provided in the diet. The number of spermatozoa per ejaculate decreased by 50% between 26 weeks and 60 weeks of age in the birds fed the maize oil diet. This age-related decrease in the number of spermatozoa was almost completely prevented by feeding the birds with the oils enriched in either 20:4n-6 or 22:6n-3. Testis mass at 60 weeks of age was approximately 1.5 times greater in birds given of the arasco and tuna orbital oil diets compared with those given the maize oil diet.     

Recycling of docosahexaenoic acid in rat retinas during n-3 fatty acid deficiency.

About 50% of the fatty acids in retinal rod outer segments is docosahexaenoic acid [22:6(n-3)], a member of the linolenic acid [18:3(n-3)] family of essential fatty acids. Dietary deprivation of n-3 fatty acids leads to only modest changes in 22:6(n-3) levels in the retina. We investigated the mechanism(s) by which the retina conserves 22:6(n-3) during n-3 fatty acid deficiency. Weanling rats were fed diets containing 10% (wt/wt) hydrogenated coconut oil (no n-3 or n-6 fatty acids), linseed oil (high n-3, low n-6), or safflower oil (high n-6, less than 0.1% n-3) for 15 weeks. The turnover of phospholipid molecular species and the turnover and recycling of 22:6(n-3) in phospholipids of the rod outer segment membranes were examined after the intravitreal injection of [2-3H]glycerol and [4,5-3H]22:6(n-3), respectively. Animals were killed on selected days, and rod outer segment membranes, liver, and plasma were taken for lipid analyses. The half-lives (days) of individual phospholipid molecular species and total phospholipid 22:6(n-3) were calculated from the slopes of the regression lines of log specific activity versus time. There were no differences in the turnover rates of phospholipid molecular species among the three dietary groups, as determined by the disappearance of labeled glycerol. Thus, 22:6(n-3) is not conserved through a reduction in phospholipid turnover in rod outer segments. However, the half-life of [4,5-3H]22:6(n-3) in the linseed oil group (19 days) was significantly less than in the coconut oil (54 days) and safflower oil (not measurable) groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Electrospray/tandem mass spectrometry for quantitative analysis of lipid remodeling in essential fatty acid deficient mice

A method utilizing electrospray ionization coupled with tandem mass spectrometry was developed as a facile and rapid method to identify and quantify lipid remodeling in vivo. Electrospray/tandem mass spectrometric analyses were performed on lipids isolated from liver tissue and resident peritoneal cells from essential fatty acid sufficient and deficient mice. Essential fatty acid deficiency was chosen as the paradigm to evaluate the methodology because it epitomizes the most extreme dietary means of altering fatty acid composition of virtually all cellular lipid species. Qualitative and quantitative changes were measured in the phospholipid and cholesterol ester species directly in the chloroform/methanol lipid extract without any prior chromatographic separation. Lipid remodeling in liver and peritoneal cells from essential fatty acid deficient mice was qualitatively similar in cholesterol ester, phosphatidylcholine, and phosphatidylethanolamine. The monoenoic fatty acids palmitoleic acid (16:1 n-7) and oleic acid (18:1 n-9) were increased markedly, whereas all n-6 and n-3 polyunsaturated fatty acids were nearly depleted in phospholipid and cholesterol ester species. The n-9 polyunsaturated fatty acid surrogate, Mead acid (20:3 n-9), substituted for arachidonic acid (20:4 n-6) and docosahexaenoic acid (22:6 n-3) in phospholipid, but not in cholesterol ester, species. Another notable difference was that adrenic acid (22:4 n-6) and docosapentaenoic acid (22:5 n-6), both metabolites of arachidonic acid, accumulated in phospholipid and cholesterol ester species of peritoneal cells, but not in liver cells, of essential fatty acid sufficient mice. The overall body of data presented illustrates the implementation of electrospray/tandem mass spectrometry as a method for facile and direct quantification of changes in lipid species during lipid metabolic studies.     

Differential effects of n-3 fatty acid deficiency on phospholipid molecular species composition in the rat hippocampus

In this study, we have examined the effects of n-3 fatty acid deficient diets on the phospholipids (PL) molecular species composition in the hippocampus. Female rats were raised for two generations on diets containing linoleic acid (18:2n-6), with or without supplementation of alpha-linolenic acid (18:3n-3) or 18:3n-3 plus docosahexaenoic acid (22:6n-3). At 84 days of age, the hippocampal phospholipids were analyzed by reversed phase HPLC-electrospray ionization mass spectrometry. Depleting n-3 fatty acids from the diet led to a reduction of 22:6n-3 molecular species in phosphatidylcholine (PC), phosphatidylethanolamine (PE), PE-plasmalogens (PLE), and phosphatidylserine (PS) by 70-80%. In general, 22:6n-3 was replaced with 22:5n-6 but the replacement at the molecular species level did not always occur in a reciprocal manner, especially in PC and PLE. In PC, the 16:0,22:6n-3 species was replaced by 16:0,22:5n-6 and 18:0,22:5n-6. In PLE, substantial increases of both 22:5n-6 and 22:4n-6 species compensated for the decreases in 22:6n-3 species in n-3 fatty acid deficient groups. While the total PL content was not affected by n-3 deficiency, the relative distribution of PS decreased by 28% with a concomitant increase in PC.The observed decrease of 22:6n-3 species along with PS reduction may represent key biochemical changes underlying losses in brain-hippocampal function associated with n-3 deficiency.     

Regiospecific distribution of fatty acids in triacylglycerols of Artemia franciscana nauplii enriched with fatty acid ethyl esters

This paper reports the positional distribution of fatty acids in triacylglycerols (TAG) of Artemia franciscana nauplii enriched with each of palmitic (16:0), oleic (18:1n-9), linoleic (18:2n-6), linolenic (18:3n-3), eicosapentaenoic (20:5n-3), and docosahexaenoic (22:6n-3) acid ethyl esters. TAG extracted from the enriched and unenriched nauplii were subjected to regiospecific analysis to determine the fatty acid compositions of the sn-1(3) and sn-2 positions of TAG. In the unenriched nauplii, 16:0, 18:1n-9, and 18:2n-6 were preferentially located in the sn-1(3) position followed by the sn-2 position [i.e. sn-1(3)>sn-2], whereas 18:3n-3 was concentrated in the sn-2 position [i.e. sn-2>sn-1(3)]. Contents of 20:5n-3 and 22:6n-3 were low. After the nauplii were enriched with each of the ethyl esters for 18 h, fatty acid fed to the nauplii showed higher content in the sn-1(3) position than in the sn-2 position [i.e. sn-1(3)>sn-2]. Distribution pattern of 18:3n-3 changed from sn-2>sn-1(3) to sn-1(3)>sn-2 during the enrichment with 18:3n-3 ethyl ester. Increases in all of the fatty acids in TAG were attributed to that in the sn-1(3) position much more than that in the sn-2 position. Artemia nauplii appear to be characterized by preferential incorporation of exogenous fatty acids into the sn-1(3) position of TAG, even though endogenous fatty acids are esterified in the opposite position.     

Captivity diets alter egg yolk lipids of a bird of prey (the American kestrel) and of a galliforme (the red-legged partridge)

The salient feature of the fatty acid profile of kestrel eggs collected in the wild was the very high proportion of arachidonic acid (15.2%+/-0.7% of fatty acid mass, n=5) in the phospholipid fraction of the yolk. Kestrels in captivity fed on day-old chickens produced eggs that differed from those of the wild birds in a number of compositional features: the proportion of linoleic acid was increased in all the lipid fractions; the proportion of arachidonic acid was increased in yolk phospholipid and cholesteryl ester; the proportion of alpha-linolenic acid was decreased in all lipid classes, and that of docosahexaenoic acid was decreased in phospholipid and cholesteryl ester. Partridge eggs from the wild contained linoleic acid as the main polyunsaturate of all the yolk lipid fractions. Captive partridges maintained on a formulated diet very rich in linoleic acid produced eggs with increased levels of linoleic, arachidonic, and n-6 docosapentaenoic acids in the phospholipid fraction; reduced proportions of alpha-linolenic acid were observed in all lipid classes, and the proportion of docosahexaenoic acid was markedly reduced in the phospholipid fraction. Thus, captive breeding of both the kestrel and the partridge increases the n-6/n-3 polyunsaturate ratio of the yolk lipids.     

The effect of n-3 fatty acids on osmotic fragility of rat erythrocytes

In order to study the effect of n-3 fatty acids on the physical state of the erythrocyte membrane, measured as osmotic fragility, rats were fed a diet supplemented in n-3 fatty acids (1.5 ml/day, 35% 20:5, 30% 22:6) for 21 days. With salt concentrations ranging from 0.37% to 0.44%, osmotic resistance was increased by 25% to 45% in cells from n-3-fed animals compared to controls. No change was observed in either phospholipid or cholesterol content in the membrane. A small, but still significant difference (P less than 0.05) in phospholipid sub-class distribution was observed in that the phosphatidylethanolamine fraction was decreased and the phosphatidylserine fraction increased after n-3 supplementation. The major change was, however, that the level of eicosapentaenoic acid (20:5(n-3] in phospholipids was increased from 1.5% of total fatty acids to 4.5%. This increase was mainly at the expense of linoleic acid (18:2(n-6]. No change was observed in the level of docosahexaenoic acid (22:6(n-3]. It is thus concluded that both the fatty acid composition and the nature of the phospholipid polar head group may influence the osmotic fragility of erythrocytes.     

Fatty acid incorporation is decreased in astrocytes cultured from alpha-synuclein gene-ablated mice

Because alpha-synuclein may function as a fatty acid binding protein, we measured fatty acid incorporation into astrocytes isolated from wild-type and alpha-synuclein gene-ablated mice. alpha-Synuclein deficiency decreased palmitic acid (16:0) incorporation 31% and arachidonic acid [20:4 (n-6)] incorporation 39%, whereas 22:6 (n-3) incorporation was unaffected. In neutral lipids, fatty acid targeting of 20:4 (n-6) and 22:6 (n-3) (docosahexaenoic acid) to the neutral lipid fraction was increased 1.7-fold and 1.6-fold, respectively, with an increase in each of the major neutral lipids. This was consistent with a 3.4- to 3.8-fold increase in cholesteryl ester and triacylglycerol mass. In the phospholipid fraction, alpha-synuclein deficiency decreased 16:0 esterification 39% and 20:4 (n-6) esterification 43% and decreased the distribution of these fatty acids, including 22:6 (n-3), into this lipid pool. alpha-Synuclein gene-ablation significantly decreased the trafficking of these fatty acids to phosphatidylinositol. This observation is consistent with changes in phospholipid fatty acid composition in the alpha-synuclein-deficient astrocytes, including decreased 22:6 (n-3) content in the four major phospholipid classes. In summary, these studies demonstrate that alpha-synuclein deficiency significantly disrupted astrocyte fatty acid uptake and trafficking, with a marked increase in fatty acid trafficking to cholesteryl esters and triacylglycerols and decreased trafficking to phospholipids, including phosphatidylinositol.     

Enrichment of platelet phospholipids with eicosapentaenoic acid and docosahexaenoic acid inhibits thromboxane A2/prostaglandin H2 receptor binding and function

Human platelet lipids were enriched in vitro with different amounts of either docosahexaenoic acid (22:6n-3), eicosapentaenoic acid (20:5n-3) or linoleic acid (18:2n-6). Of the total fatty acid incorporated, between 82 and 95% was associated with the phospholipid (PL) fraction, with the remainder as either neutral lipid or hydroxy fatty acid. Within the PL fraction, the majority (64% of total) of each fatty acid was incorporated into phosphatidylcholine. It was found that platelet aggregation induced by the thromboxane A2/prostaglandin H2 mimetic (15S)-hydroxy-11,9-(epoxymethano)prosta-5Z,13E-dienoic acid (U46619) was inhibited after PL enrichment with 22:6n-3 or 20:5n-3, but not after 18:2n-6 enrichment. The specificity of 22:6n-3 and 20:5n-3 for U46619 activation was demonstrated by the finding that neither fatty acid significantly inhibited thromboxane A2/prostaglandin H2-independent aggregation induced by A23187 or thrombin. Furthermore, enrichment with 22:6n-3 or 20:5n-3 resulted in inhibition of [3H]U46619 specific binding, while enrichment with 18:2n-6 did not affect binding. Scatchard analysis revealed that thromboxane A2/prostaglandin H2 receptor affinity for [3H]U46619 decreased 4.8-fold following 22:6n-3 incorporation. These results demonstrate that platelet phospholipid enrichment with 22:6n-3 or 20:5n-3 results in a selective inhibition of thromboxane A2/prostaglandin H2 receptor function.     

The relationship between the fatty acid profiles of the yolk and the embryonic tissue lipids: A comparison between the lesser black backed gull (Larus fuscus) and the pheasant (Phasianus colchicus)

Although substantial information is available regarding the fatty acid composition of lipids of the yolk and of the developing tissues of the chicken embryo, there is little knowledge on this topic for other avian species. The aim of the present study was to compare the yolk and embryonic tissue fatty acid profiles for a species selecting its food in the wild (the lesser black backed gull) with one fed on a standard commercial diet (the commercially reared pheasant). The fatty acid compositions of the yolk lipids were determined, and major differences were observed between the two species. In particular, the phospholipid of the gull yolk was enriched in 20:4n-6 and 22:6n-3 (18.8 and 7.1%, respectively, by weight of total fatty acids) in comparison with the pheasant (4.0 and 4.1%, respectively). The fatty acid compositions of the embryonic tissues were determined using eggs incubated in the laboratory. For the liver and heart, the fatty acid composition of the lipids in the two species reflected the initial yolk composition, with the gull tissue lipids generally containing higher proportions of 20:4n-6 and 22:6n-3 than those of the pheasant. In contrast, the fatty acid profiles of the brain phospholipid were essentially identical in the two species, with 20:4n-6 and 22:6n-3 comprising approximately 9 and 17%, respectively, of total fatty acids in both cases.     

Phospholipid fatty acid composition of pulmonary airway epithelial cells: potential substrates for oxygenation

To determine possible substrates for airway epithelial lipoxygenase and cyclooxygenase activities, we examined the amounts and distributions of fatty acids in phospholipids of human, dog and sheep airway epithelial cells. We found that the cells contained significant levels of n-6 and n-3 fatty acids in species-specific amounts: dog cells were relatively enriched in the n-6 series and poor in n-3, while sheep cells were enriched in the n-3 series and poor in n-6. Despite differences in fatty acid content, cells from each species expressed a constant phospholipid composition and distributed their n-6 and n-3 fatty acids in a stereotyped fashion among phospholipid classes. The analysis shows that the species differences in oxygenation activities reported previously are paralleled by heterogeneity in potential substrates.     

Changes in fatty acid composition of Mytilus galloprovincialis (Lmk) fed on microalgal and wheat germ diets

Dietary fatty acid incorporation and changes in various lipid and phospholipid classes in the mussel Mytilus galloprovincialis subjected to three different dietary regimens were analysed and compared. Group A was unfed; group B received a diet consisting of 100% Thalassiosira weissflogii, exhibiting the typical fatty acid composition of diatoms, and group C received a diet consisting of 100% wheat germ conferring a 18:2:n-6 abundance. Biochemical analyses of diets and mussels were carried out at the beginning and at the end of the 30-day experimental period. Starvation and T. weissflogii based diet poorly affected mussel growth and fatty acid composition which remained unchanged. On the contrary, the wheat germ-based diet increased the condition index and deeply affected the fatty acid profile of all lipid and phospholipid classes. The high dietary 18:2n-6 level drastically reduced tissue content of 20:4n-6, 20:5n-3 and 22:6n-3. The biosynthesis of Non Methylene Interrupted (NMI) dienoic fatty acid appeared to be insensitive to the high input of 16:1n-7 and 18:1n-9 respectively from diet B and C, and to the PUFA shortage of diet C. Nevertheless the two NMI trienoic derivatives, 20:3Δ5,11,14 and 22:3Δ7,13 16, were found higher in C with respect to other groups, presumably due to the high 18:2n-6 content of this diet.     

Maternal dietary fat alters amniotic fluid and fetal intestinal membrane essential n-6 and n-3 fatty acids in the rat

We investigated whether maternal fat intake alters amniotic fluid and fetal intestine phospholipid n-6 and n-3 fatty acids. Female rats were fed a 20% by weight diet from fat with 20% linoleic acid (LA; 18:2n-6) and 8% alpha-linolenic acid (ALA; 18:3n-3) (control diet, n = 8) or 72% LA and 0.2% ALA (n-3 deficient diet, n = 7) from 2 wk before and then throughout gestation. Amniotic fluid and fetal intestine phospholipid fatty acids were analyzed at day 19 gestation using HPLC and gas-liquid chromotography. Amniotic fluid had significantly lower docosahexaenoic acid (DHA; 22:6n-3) and higher docosapentaenoic acid (DPA; 22:5n-6) levels in the n-3-deficient group than in the control group (DHA: 1.29 +/- 0.10 and 6.29 +/- 0.33 g/100 g fatty acid; DPA: 4.01 +/- 0.35 and 0.73 +/- 0.15 g/100 g fatty acid, respectively); these differences in DHA and DPA were present in amniotic fluid cholesterol esters and phosphatidylcholine (PC). Fetal intestines in the n-3-deficient group had significantly higher LA, arachidonic acid (20:4n-6), and DPA levels; lower eicosapentaenoic acid (EPA; 20:5n-3) and DHA levels in PC; and significantly higher DPA and lower EPA and DHA levels in phosphatidylethanolamine (PE) than in the control group; the n-6-to-n-3 fatty acid ratio was 4.9 +/- 0.2 and 32.2 +/- 2.1 in PC and 2.4 +/- 0.03 and 17.1 +/- 0.21 in PE in n-3-deficient and control group intestines, respectively. We demonstrate that maternal dietary fat influences amniotic fluid and fetal intestinal membrane structural lipid essential fatty acids. Maternal dietary fat can influence tissue composition by manipulation of amniotic fluid that is swallowed by the fetus or by transport across the placenta.     

Sex differences in the metabolism of essential fatty acids studied in isolated rat liver cells

The triacylglycerol synthesis from exogenous linoleic acid (18:2(n-6], linolenic acid (18:3(n-3], dihomogammalinolenic acid (20:3(n-6], eicosapentaenoic acid (20:5(n-3] and oleic acid (18:1(n-9] was observed to be significantly increased in isolated liver cells from female rats compared with males. The rate of fatty acid oxidation and phospholipid biosynthesis was concomitantly more important in male cells. With the C22-polyenoic fatty acids, adrenic acid (22:4(n-6] and docosahexaenoic acid (22:6(n-3), only a minor sex-related difference in fatty acid metabolism was found.     

Phosphatidylethanolamine methyltransferase: evidence for influence of diet fat on selectivity of substrate for methylation in rat brain synaptic plasma membranes

Male weanling rats were fed diets containing 20% (w/w) fat differing in fatty acid composition for 24 days. Synaptic plasma membranes were isolated from the brain and the fatty acid composition of phosphatidylethanolamine and phosphatidylcholine was determined. In vitro assays of phosphatidylethanolamine methyl-transferase activity were performed on fresh membrane samples to assess effect of dietary fat on the rate of phosphatidylethanolamine methylation for phosphatidylcholine synthesis via the phosphatidylethanolamine methyltransferase pathway. Dietary level of n-6 and ratio of n-6 to n-3 fatty acids influenced membrane phospholipid fatty acid composition and activity of the lipid-dependent phosphatidylethanolamine methyltransferase pathway. Rats fed a diet rich in n-6 fatty acids produced a high ratio of n-6/n-3 fatty acids in synaptosomal membrane phosphatidylethanolamine, and elevated rates of methylation of phosphatidylethanolamine to phosphatidylcholine by phosphatidylethanolamine methyltransferases, suggesting that the pathway exhibits substrate selectivity for individual species of phosphatidylethanolamine containing long-chain homologues of dietary n-6 and n-3 fatty acids (20:4(n-6), 22:4(n-6), 22:5(n-6) and 22:6(n-3). It may be concluded that diet alters the membrane content of n-6, n-3 and monounsaturated fatty acids, and that change in phosphatidylethanolamine species available for methylation to phosphatidylcholine alters the rate of product synthesis in vivo by the phosphatidylethanolamine methyltransferase pathway.     

Selective incorporation of various C-22 polyunsaturated fatty acids in Ehrlich ascites tumor cells

Three 14C-labeled 22-carbon polyunsaturated fatty acids, 7,10,13,16-[14C]docosatetraenoic acid (22:4(n-6)), 7,10,13,16,19-[14C]docosapentaenoic acid (22:5(n-3)), and 4,7,10,13,16,19-[14C]docosahexaenoic acid (22:6(n-3)), were compared with [3H]arachidonic acid (20:4(n-6] and [14C]linoleic acid (18:2(n-6)) to characterize their incorporation into the lipids of Ehrlich ascites cells. The relatively rapid incorporation of the labeled 22-carbon acids into phosphatidic acid indicated that substantial amounts of these acids may be incorporated through the de novo pathway of phospholipid synthesis. In marked contrast to 20:4(n-6), the 22-carbon acids were incorporated much less into choline glycerophospholipids (CGP) and inositol glycerophospholipids (IGP). No selective preference was apparent for the (n-3) or (n-6) type of fatty acids. The amounts of the acids incorporated into diacylglycerophosphoethanolamine were in the order of: 22:6(n-3) greater than 20:4(n-6) much greater than 22:5(n-3) greater than or equal to 22:4(n-6) greater than 18:2(n-6), whereas for alkylacylglycerophosphoethanolamine they were in the order of: 22:4(n-6) greater than 22:6(n-3) greater than 22:5(n-3) much greater than 20:4(n-6) greater than 18:2(n-6). Of the mechanisms possibly responsible for the selective entry of 22-carbon acids into ethanolamine glycerophospholipids, the most reasonable explanation was that the cytidine-mediated ethanolamine phosphotransferase may have a unique double selectivity: for hexaenoic species of diacylglycerol and for 22-carbon polyunsaturated fatty acid-containing species of alkylacylglycerol. The relative distribution of fatty acids between newly incorporated and already maintained lipid classes suggested that IGP may function in Ehrlich cells as an intermediate pool for the retention of polyunsaturated fatty acids in glycerolipids.     

Fatty acids specifically related to the anisotropic properties of plasma membrane from rat urothelium

Four different luminal surfaces of rat urothelium differing in their fatty acid composition were prepared by dietary induction. In order to induce lipid changes, each of four groups of rat received a basal diet rich in one of the unsaturated n-3, n-6 or n-9 fatty acid families and a commercial (control) diet. The effects of the dietary regime on the fatty acid composition of luminal urothelial membranes and their relation to the mobility of fluorescent probes were studied. In comparison with the control diet membrane, all three fatty acid-rich diets induced a decrease of the percentage amount of saturated fatty acid while that of the unsaturated fatty acids was increased. Accordingly, all three diets increased the unsaturation index in comparison with the control diet. The anisotropy across each membrane fraction was assessed using the n-(9-anthroyloxy) fatty acid fluorescent probes 3-AS, 7-AS and 12-AS, which locate at different depths in the membrane. Two different anisotropy profiles were observed. One profile showed the highest anisotropy at the C7 depth, whereas the other exhibited a continuous decrease of the anisotropy from the surface to the center of the bilayer. The molecular properties (isomerization) of 18:2n-9 fatty acid may account, at least in part, for the observed V-shaped profile (the ascending trend) of the membrane anisotropy values as a function of the respective 18:2n-9 fatty acid contents. Nevertheless, the minimum value of the profile did not correspond to the minimum 18:2n-9 fatty acid content, but rather to the higher amount of docosahexaenoic (22:6n-3) fatty acid. Thus, a modulating role of the 22:6n-3 fatty acid on the rigidifying effect of 18:2n-9 fatty acid is suggested, possibly mediated by relationships between fatty acid composition, saturated and unsaturated chain lengths, and freedom of motion of the phospholipid acyl chains.     

Lipid and fatty acid composition during embryo and larval development of puye Galaxias maculatus Jenyns, 1842, obtained from estuarine, freshwater and cultured populations

Galaxias maculatus eggs and larvae obtained from broodfish captured either in an estuarine or a freshwater environment, as well as from cultured broodstock were analysed to compare their lipid and fatty acid profiles. Results showed a lower lipid content in embryos and larvae from estuarine populations than those from fresh water, denoting the influence of environmental conditions. The n-3:n-6 ratio was higher in eggs from estuarine and cultured populations, being in the range of marine fishes, whereas for eggs from freshwater fish was lower and typical of freshwater fishes. The polyunsaturated fatty acids (PUFA), particularly docosahexaenoic acid (22:6n-3) and eicosapentaenoic acid (20:5n-3), were higher in eggs and larvae of broodstock coming from culture or estuarine environments than in those from fresh water. Moreover, these fatty acids markedly increased after hatching in larvae coming from estuarine populations, suggesting the effect of the environment on fatty acid profiles to physiologically prepare the larvae to adapt to higher salinity conditions. Linoleic acid (18:2n-6) content was higher in fresh water fish and its reduction during embryo and larval development was accompanied by a significant increase of arachidonic acid (20:4n-6), which was not observed in embryos or larvae from broodstock fish from estuary or aquaculture origin. Both environment and diet of broodstock fish affected lipid and fatty acid composition of G. maculatus embryo and larvae as well as their changes during development.