Involvement of phospholipid molecular species in controlling structural order of vertebrate brain synaptic membranes during thermal evolution

Fluorescence anisotropy parameter of [p-(6-phenyl)-1,3,5-hexatrienyl]phenyl-propionic acid (DPH-PA) and 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) embedded in synaptic plasma membranes prepared from brains of cold (5°C) and warm (22°C) adapted fish (Cyprinus carpio L.), rat (Rattus norvegicus) and bird (Branta canadensis), was studied. Fatty acid composition of total lipids as well as molecular species composition of diacyl phosphatidylcholines and phosphatidylethanolamines was also determined. The amount of long-chain polyunsaturated fatty acids decreased with increasing body temperature. There was a nearcomplete compensation of membrane structural order for environmental/body tempeature over the evolutionary scale as seen by DPH-PA. Using TMA-DPH, the compensation was partial with rat and bird. Since DPH-PA and TMA-DPH differ in their charges, it is proposed, that the former reported membrane regions rich in cationic or zwitterionic (neutral) phospholipids and the latter, membrane regions rich in negatively charged phospholipids in the synaptic plasma membranes. Many different molecular species (20–25) of diacyl phosphatidylcholines and diacyl phosphatidylethanolamines were identified. The level of 16:0/22:6 phosphatidylcholine decreased while disaturated phosphatidylcholines increased with increase of environmental/body temperature from the fish through the bird. Level of 1-monoenoic 2-polyenoic phosphatidylethanolamines also decreased with an increase in environmental/body temperature. Experiments using vesicles made of mixed synthetic phosphatidylcholine vesicles (16:0/16:0, 16:0/18:1, 16:0/22:6 in various proportions) showed that increase in disaturated phosphatidylcholine species does not explain the observed complete adjustment of membrane structural order in synaptic plasma membranes. Change in level of 1-monoenoic, 2-polyenoic phosphatidylethanolamines might be one of the factors involved in controlling the biophysical properties of the membrane according to the temperature.     

Dietary linoleic acid and the fatty acid profiles in rats fed partially hydrogenated marine oils

The influence of the linoleic acid levels of diets containing partially hydrogenated marine, oils (HMO) rich in isomeric 16∶1, 18∶1, 20∶1 and 22∶1 fatty acids on the fatty acid profiles of lipids from rat liver, heart and adipose tissue was examined. Five groups of rats were fed diets containing 20 wt% fat−16% HMO+4% vegetable oils. In these diets, the linoleic acid contents varied between 1.9% and 14.5% of the dietary fatty acids, whereas the contents oftrans fatty acids were 33% in all groups. A sixth group was fed a partially hydrogenated soybean oil (HSOY) diet containing 8% linoleic acid plus 32%trans fatty acids, mainly 18∶1, and a seventh group, 20% palm oil (PALM), with 10% linoleic acid and notrans fatty acids. As the level of linoleic acid in the HMO diets increased from 1.9% to 8.2%, the contents of (n−6) polyunsaturated fatty acids (PUFA) in the phospholipids increased correspondingly. At this dietary level of linoleic acid, a plateau in (n−6) PUFA was reached that was not affected by further increase in dietary 18∶2(n−6) up to 14.5%. Compared with the HSOY- or PALM-fed rats, the plateau value of 20∶4(n−6) were considerably lower and the contents of 18∶2(n−6) higher in liver phosphatidylcholines (PC) and heart PC. Heart phosphatidylethanolamines (PE) on the contrary, had elevated contents of 20∶4(n−6), but decreased 22∶5(n−6) compared with the PALM group. All groups fed HMO had similar contents oftrans fatty acids, mainly 16∶1 and 18∶1, in their phospholipids, irrespective of the dietary 18∶2 levels, and these contents were lower than in the HSOY group. High levels of linoleic acid consistently found in triglycerides of liver, heart and adipose tissue of rats fed HMO indicated that feeding HMO resulted in a reduction of the conversion of linoleic acid into long chain PUFA that could not be overcome by increasing the dietary level of linoleic acid.     

Fatty chain compositon of ether and ester glycerophospholipids in the Japanese oysterCrassostrea gigas (Thunberg)

The fatty chain compositions of 1-O-alk-1′-enyl-2-acyl, 1-0-alkyl-2-acyl, and 1,2-diacyl glycerophospholipids of the Japanese oysterCrassostrea gigas (Thunberg) were investigated. Major fatty chains in thesn-1 position of 1-alk-1′-enyl-2-acyl ethanolamine phospholipids (EPL) were 18∶0 (64.7%) and 20∶1 (11.1%). Majorsn-1 chains of alkenylacyl choline phospholipids (CPL) were 18∶0 (63.3%) and 16∶0 (22.2%). In the case of 1-alkyl-2-acyl EPL, the predominant fatty chains in thesn-1 position were 18∶0 (51.5%), 16∶0 (16.0%) and 20∶1 (12.5%); in the case of 1-alkyl-2-acyl CPL, the majorsn-1 chains were 16∶0 (44.0%) and 14∶0 (23.4%). Saturated fatty chains were predominant in both EPL and CPL. Prominent fatty acids in thesn-2 position of the alkenylacyl EPL were 22∶6n−3 (29.0%), 20∶5n−3 (19.0%) and 22∶2 NMID (non-methylene interrupted dienes, 16.6%) contributing to about 65% of the total fatty acids, while alkenylacyl CPL was rich in the saturated acids 16∶0 (32.0%) and 18∶0 (9.2%). In the alkylacyl EPL, 16∶0, 18∶1n−9, 18∶0 and 16∶1n−7 were prominentsn-2 fatty acids and accounted for 30.6%, 10.0%, 9.8%, and 8.3%, respectively. Polyunsaturated fatty acids were detected, but were present at extremely low percentages. Majorsn-2 fatty acids in alkylacyl CPL were 16∶0 (25.4%), 22∶6n−3 (16.0%) and 20∶5n−3 (8.4%). The major fatty acids of diacyl EPL were 20∶5n−3 (22.3%), 16∶0 (17.9%), and 18∶0 (16.1%), and those of diacyl CPL were 16∶0 (30.4%), 20∶5n−3 (17.6%) and 18∶1n−7 (7.4%).     

Fatty acid composition of brain phospholipids in aging and in Alzheimer’s disease

The two major phospholipid classes, namely, phosphatidylethanolamines (PE) and phosphatidylcholines (PC), were studied in four different regions of human brain,i.e., in frontal gray matter, frontal white matter, hippocampus and in pons. The fatty acid (FA) compositions of these phospholipids were found to be specific for the different regions. PC contains mostly saturated and 18∶1 FA, while PE is rich in polyunsaturated FA. Aging has no influence on the FA compositions, while in Alzheimer’s disease (AD) PE is modified in all four regions, particularly in frontal gray matter and in hippocampus. The abundance of the major monounsaturated FA of PE, 18∶1, is not significantly altered in Alzheimer’s disease, but there is a substantial increase in the relative amounts of the saturated components 14∶0, 16∶0 and 18∶0. This is paralleled by a decrease in the polyunsaturated FA 20∶4, 22∶4 and 22∶6. It is not clear whether the changes observed are specific for AD. Changes in saturated/polyunsaturated FA ratio are likely to influence cellular function, which in turn may cause certain neural deficiencies. The findings do not support the hypothesis that AD reflects an accelerated aging process.     

Effect of pH on the affinity of phospholipids for cholesterol

The ability of multilamellar vesicles of phosphatidylcholine and phosphatidylethanolamine in aqueous phase to prevent access to cholesterol by a nonpolar solvent was examined. Phosphatidylethanolamine vesicles retained less sterol than phosphatidylcholine vesicles. In mixed vesicles, cholesterol was retained in proportion to the amount of phosphatidylcholine. To alter the charge and hydration of head groups, pH was adjusted from 1.2 to 12.5. Above pH 8, both phosphatidylethanolamine and phosphatidylcholine retained sterol in a 1∶1 molar ratio of phospholipid to cholesterol, regardless of acyl side chain composition. Between pH 2.0 and pH 8.0, sterol retention varied with type of head group and side chain. Lipids with 16-carbon saturated side chains retained more sterol than 18-carbon unsaturated or 12-carbon saturated side chains. Between pH 1.1 and 2.0, none of the phosphatidylethanolamines retained sterol, but long chain phosphatidylcholines, saturated or unsaturated, retained sterol in a 1∶1 molar ratio of phospholipid to sterol. Short chain phosphatidylethanolamines and phosphatidylcholines retained 0 to 20% at the low- to mid-pH range. Size of multilamellar vesicles, measured by Doppler effect light scattering analysis, had no bearing on sterol retention. Sonication of vesicles, which increases surface curvature, increases the retention of sterol. Fluorescence polarization indicated that cholesterol does not interact with DPPC or DLPC side chains. The observations can be interpreted in terms of space requirements of head groups, including charge repulsion and hydration. Other factors, such as monovalent cation replacement by protons, juxtaposition of charged groups on vesicle surfaces and length and unsaturation of acyl side chains affect the affinity of phospholipids for cholesterol.     

Molecular species composition of the major diacyl glycerophospholipids from muscle,liver, retina and brain of cod (Gadus morhua)

The molecular species composition of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS) from white muscle, liver, retina and brain of cod (Gadus morhua) were determined by high-performance liquid chromatography of the respective 1,2-diacylglycerol 3,5-dinitrobenzoyl derivatives. A minimum of 69 diacyl species was identified. In muscle and liver saturated fatty acid/polyunsaturated fatty acid (PUFA) and monounsaturated fatty acid/PUFA molecular species were predominant, particularly 16∶0/20∶5 and 16∶0/22∶6 in PC, 16∶0/22∶6 and 18∶1/22∶6 in PE and 18∶0/22∶6 and 18∶1/22∶6 in PS. Didocosahexaenoyl species were major components of PC, PE and PS from retina, comprising 29.3, 71.8 and 59.7% of the respective totals. Didocosahexaenoyl species were also abundant in PE and PS from brain, accounting for 13.8 and 24.0% of the totals, respectively. DiPUFA species were important in muscle, totalling 21.2% in PC and 38.3% in PE. PC from all tissues had the largest amounts of species containing only saturated or monounsaturated fatty acids, accounting for 59.8% of PC from brain, including 12.8% of 18∶1/24∶1 plus 24∶1/18∶1.     

Fatty acid composition of subcellular particles from sheep platelets and topological distribution of phosphatidylethanolamine fatty acids in the plasma membrane

The fatty acid composition of individual phospholipids in subcellular fractions of sheep platelets and the asymmetrical distribution of phosphatidylethanolamine (PE) fatty acyl chains across the plasma membrane were examined. The main fatty acids of total lipid extracts were oleic (18∶1; 32–41%), linoleic (18∶2, 10–17%), stearic (18∶0; 13–15%), palmitic (16∶0; 11–15%) and arachidonic (20∶4; 8–12%) acids, with a saturated/unsaturated ratio of about 0.4. Each phospholipid class had a distinct fatty acid pattern. Sphingomyelin (SM) showed the highest degree of saturation (50%), with large proportions of behenic (22∶0), 18∶0 and 16∶0 acids. The main fatty acid in PE, phosphatidylserine (PS) and phosphatidylcholine (PC) was 18∶1n−9. Our findings suggest that fatty acids are asymmetrically distributed between thecholineversus the non-choline phospholipids, and also between plasma membranes and intracellular membranes. The transbilayer distribution of PE fatty acids in plasma membranes from non-stimulated sheep platelets was investigated using trinitrobenzenesulfonic acid (TNBS). A significant degree of asymmetry was found, which is a new observation in a non-polar cell. The PE molecules from the inner monolayer contained higher amounts of 18∶2 and significantly less 18∶1 and 20∶5 than those found in the outer monolayer, although no major differences were detected in the transbilayer distribution of total unsaturatedversus saturated PE acyl chains.     

Molecular speciation of fish sperm phospholipids: Large amounts of dipolyunsaturated phosphatidylserine

The molecular species compositions of the main diacyl phosphoglyceride classes and ether-linked subclasses from sperm of three species of fish, sea bass Dicentrarchus labrax, Atlantic salmon Salmo salar and Chinook salmon Onchorhynchus tsawytscha, were determined. The phospholipids from sperm were highly unsaturated, dipolyunsaturated fatty acid (diPUFA) molecular species comprised 64.6 to 71.8% of phosphatidylserine (PS), 10.1 to 17.4% of phosphatidylethanolamine (PE), and 3.3 to 10.1% of phosphatidylcholine (PC). In sea bass sperm, di22∶6n-3 phospholipid was the predominant diPUFA molecular species, but in both salmon species 22∶5n-3/22∶6n-3 was also a major constituent of PS. Phospholipids containing 22∶6n-3 dominated in sea bass sperm with 16∶0/22∶6n-3 as a major component of PC and PE, and 18∶0/22∶6n-3 of PE and PS in addition to di22∶6n-3 in the latter two classes. In contrast, both salmon species contained much more 20∶5n-3 and less 22∶6n-3 so that saturated/20∶5n-3 and monounsaturated/20∶5n-3 molecular species were more abundant than the corresponding molecules containing 22∶6n-3. Ether-linked lipids comprised 11.3–36.3% of choline and ethanolamine phosphoglycerides in each fish species. Molecular species containing 22∶6n-3 were the major components of 1-O-alkyl-2-acyl-glycerophosphocholine, especially 16∶0a/22∶6n-3 in sea bass and 18∶1a/∶6n-3 in the two salmon species, while in 1-O-alk-1′-enyl-2-acyl-glycerophosphoethanolamine, 16∶0a/22∶6n-3 was the major component in both salmon and 18∶0a/22∶6n-3 in sea bass with 18∶1a/22∶6n-3 abundant in all three species. In Atlantic salmon 1-O-alkyl-2-acylglycerophosphoethanolamine comprised 24.6% of ethanolamine glycerophospholipids which were predominantly 16∶0a/22∶6n-3 and 18∶1a/22∶6n-3. Phosphatidylinositol from sperm was dominated by stearoyl/C₂₀ PUFA molecular species, in sea bass overwhelmingly 18∶0/20∶4n-6, while in both salmon species 18∶0/20∶4n-6 and 18∶0/20∶5n-3 were equally abundant.     

Molecular species composition of glycerophospholipids from white matter of human brain

The molecular species composition of the major glycerophospholipids from white matter of human brain were determined by high-performance liquid chromatography of the 3,5-dinitrobenzoyl derivatives of the corresponding diradylglycerols. In phosphatidylcholine (PC) and phosphatidylserine (PS), molecular species containing only saturated fatty acids (SFA) and monounsaturated fatty acids (MUFA) comprised 85.7 and 82.4% of the respective totals, with 18∶0/18∶1 predominant in PS and 16∶0/18∶1 in PC. These molecular species were also abundant in phosphatidylethanolamine (PE), but in this phospholipid species containing polyunsaturated fatty acids (PUFA), largely 18∶0/22∶6n−3 and 18∶0/20∶4n−6, accounted for over half the total; 18∶1/18∶1 was also abundant in PE. In contrast, 1-O-alk-1′-enyl-2-acylsn-glycero-3-phosphoethanolamine (GPE) had much more SFA- and MUFA-containing species, predominantly 16∶0a/18∶1, 18∶0a/18∶1 and 18∶1a/18∶1, with low amounts of species containing 20∶4n−6 and 22∶6n−3. In alkenylacyl GPE, 22∶4n−6 was the major PUFA and 16∶0a/22∶4n−6 and 18∶1a/22∶4n−6 the main PUFA-containing species. There was six times more 22∶6n−3, twice as much 20∶4n−6 and half the amount of 22∶4n−6 in PE as compared to alkenylacyl GPE. Molecular species are abbreviated as follows:e.g., 16∶0/18∶1 PE is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine; the corresponding alkenylacyl species, 1-O-hexadec-1′-enyl-2-oleoyl-sn-glycero-3-phosphoethanolamine is 16∶0a/18∶1.     

Phospholipid molecular species composition of developing fetal guinea pig brain

Adequate accumulation of polyunsaturated essential fatty acids, in particular docosahexaenoic acid (22∶6n−3), into membrane phospholipids is critical for optimal fetal brain development. This process is maximal during the period of rapid neurite outgrowth, neuritogenesis, which precedes the major growth phase, myelination. There is no information about differential changes during gestation to individual brain phospholipid molecular species which contain 22∶6n−3. Such details of brain development would be concealed by total fatty acid analysis of isolated phospholipid classes. We have detailed phosphatidylcholine (PC) and phosphatidylethanolamine (PE) molecular species compositions in developing fetal guinea pig brain. Total brain PC concentration increased substantially between 40 and 68 (term) d of gestation, corresponding to myelination, while PE increased in a biphasic manner between 25–35 d, which was coincident with onset of neuritogenesis, and 40–68 d. Fetal brain development was accompanied by complex changes in the concentration of individual phospholipid molecular species. During early gestation (25–40 d) 22∶6n−3 was enriched in both PC and PEsn−1 16∶0 molecular species. However, between 40 d and term there was no further increase in brain PC 22∶6n−3 content, while brain PE was significantly enriched in both PE 18∶1/22∶6 and PE18∶0/22∶6. We hypothesize that accumulation of 22∶6n−3 intosn−1 18∶1 and 18∶0 species represents establishment of a 22∶6n−3-containing membrane PE pool which may be turned over more slowly thansn−1 16∶0 species. Identification of specific changes in membrane phospholipids which are associated with defined events in brain development may provide a basis for assigning functional roles to individual molecular species.     

Positional analysis of triglycerides and phospholipids rich in long-chain polyunsaturated fatty acids

Four sources of long-chain polyunsaturated fatty acids (LCP) differing in their chemical structure (triglycerides or phospholipids) and in their origin (tuna triglycerides, fungal triglycerides, egg phospholipids, and pig brain phospholipids) were analyzed to determine the distribution of the component fatty acids within the molecule. Lipase and phospholipase A₂ hydrolysis was performed to obtain 2-monoacylglycerols and lysophospholipids, respectively, which allowed us to determine the distribution of fatty acids between the sn-2 and sn-1,3 positions of triglycerides or between the sn-1 and sn-2 position of phospholipids. Fatty acids in the LCP sources analyzed were not randomly distributed. In tuna triglycerides, half of the total amount of 22∶6n−3 was located at the sn-2 position (49.52%). In fungal triglycerides, 16∶0 and 18∶0 were esterified to the sn-1,3 (92.22% and 91.91%, respectively) 18∶1 and 18∶2 to the sn-2 position (59.77% and 62.62%, respectively), and 45% of 20∶3n−6 and only 21.64% of 20∶4n−6 were found at the sn-2 position. In the lipid sources containing phospholipids, LCP were mainly esterified to the phosphatidylethanolamine fraction. In egg phospholipids, most of 20∶4n−6 (5.50%, sn-2 vs. 0.91%, sn-1) and 22∶6n−3 (2.89 vs. 0.28%) were located at the sn-2 position. In pig brain phospholipids, 22∶6n−3 was also esterified to the sn-2 (13.20 vs. 0.27%), whereas 20∶4n−6 was distributed between the two positions (12.35 vs. 5.86%). These results show a different fatty acid composition and distribution of dietary LCP sources, which may affect the absorption, distribution, and tissue uptake of LCP, and should be taken into account when supplementing infant formulas.     

Phospholipid molecular species from human placenta lipids

The phospholipid molecular species from a large-scale preparation of human placenta lipids were analyzed. The major placental phospholipids were choline glycerophospholipids (CPL) (53.2 wt%), sphingomyelin (21.7 wt%) and ethanolamine glycerophospholipids (EPL) (14.6 wt%). 1,2-Diacyl-glycerophosphocholine was the most abundant subclass of CPL (91.7 mol%), while EPL contained 1,2-diacyl (54.6 mol%) and 1-alk-1′-enyl-2-acyl (43.8 mol%) subclasses. The level of polyunsaturated fatty acids (PUFA) in total phospholipids was remarkably constant (38.4–39.9 mol%) within all placental batches tested. The long-chain PUFA, mainly 20∶4n−6 and 22∶6n−3 of the n−6 and n−3 series, respectively, were found in high proportion in all phospholipid classes, especially in EPL (46.7 mol%) and in inositol glycerophospholipids (IPL) (39.9 mol%). CPL and serine glycerophospholipids were much richer in 18∶1n−9 and 18∶2n−6. High levels of molecular species with arachidonic acid in thesn-2 position were found particularly in 1-alk-1′-enyl-2-acyl-glycerophospho-ethanolamine (with 24.0 mol% 16∶0 and 22.0 mol% 18∶0 insn-1 position) and in 1,2-diacyl glycerophosphoinositol with 42.6 mol% 18∶0 insn-1 position. EPL subclasses were rich in 22∶6n−3, which occurs mainly as 16∶0/22∶6n−3 (11.7 mol%) in the polasmalogen form and as 18∶0/22∶6n−3, 16∶0/22∶6n−3 and 18∶1/22∶6n−3 in the diacyl forms. Based on their availability and composition, placental phospholipids could be of interest, for example, for supplementing artificial milk preparations with n−3 and n−6 long-chain PUFA for newborn infants with insufficiently developed 18∶2n−6 and 18∶3n−3 desaturation/elongation.     

Incorporation ofcis-octadecenoic acids into the rat liver mitochondrial membrane phospholipids and adipose tissue triglycerides

The incorporation of the dietarycis 18∶1 (n−12) andcis 18∶1 (n−10) into liver mitochondrial membrane phospholipids and adipose tissue trigly cerides was studied in 4 groups of rats fed diets containing 10 weight percent (wt%) of fat with the following contents of octadecenoic acids: 50%cis 18∶1(n−12) +9%cis 18∶1 (n−9); 25%cis 18∶1 (n−12)+32%cis 18∶1 (n−9); 50%cis 18∶1 (n−10)+10%cis 18∶1 (n−9); or 54%cis 18∶1 (n−9). Dietary linoleic acid was 3 wt% in all 4 groups. In the mitochondrial membranes, the isomeric octadecenoic acids were primarily incorporated into the 1-position of phosphatidylcholines and phosphatidylethanolamines at the expense of saturated fatty acids. The maximal incorporations observed in the 1-position of phosphatidylethanolamines were 4.8% 18∶1 (n−12) and 8.9% 18∶1 (n−10). No effects on the contents of polyunsaturated fatty acids in the phospholipids were seen. In the adipose tissue, the isomeric octadecenoic acids were incorporated at a level of 13%cis 18∶1 (n−12) or 23%cis 18∶1 (n−10), paralleled by a reduction in the content of oleic acid. Presented in part at the 9th Scandinavian Symposium on Lipids, Visby, Sweden, June 1977.     

Lipid composition of the pineal organ from rainbow trout (Oncorhynchus mykiss)

The lipid composition of the pineal organ from the rainbow trout (Oncorhynchus mykiss) was determined to establish whether the involvement of this organ in the control of circadian rhythms is reflected by specific adaptations of lipid composition. Lipid comprised 4.9% of the tissue wet weight and triacylglycerols were the major lipid class present (47% of total lipid). Phosphatidylcholine (PC) was the principal polar lipid, and smaller proportions of other phospholipids and cholesterol were also present. Plasmalogens contributed 11% of the ethanolamine glycerophospholipids (EGP). No cerebrosides were detected. The fatty acid composition of triacylglycerols was generally similar to that of total lipids in which saturated, monounsaturated and polyunsaturated fatty acids (PUFA) were present in almost equal proportions. Each of the polar lipid classes had a specific fatty acid composition. With the exception of phosphatidylinositol (PI), in which 20∶4n−6 comprised 27.4% of the total fatty acids, 22∶6n−3 was the principal PUFA in all lipid classes. The proportion of 20∶5n−3 never exceeded 6.0% of the fatty acids in any lipid class. The predominant molecular species of PC were 16∶0/22∶6n−3 and 16∶0/18∶1, which accounted for 33.2 and 28.5%, respectively, of the total molecular species of this phospholipid. Phosphatidylethanolamine (PE) contained the highest level of di-22∶6n−3 (13.0%) of any phospholipid. There was also 4.9% of this molecular species in phosphatidylserine (PS) and 4.1% in PC. In PE, the species 16∶0/22∶6, 18∶1/22∶6 and 18∶0/22∶6 totalled 45.1%, while in PS 18∶0/22∶6 accounted for 43.9% of the total molecular species. The most abundant molecular species of PI was 18∶0/20∶4n−6 (37.8%). The lipid composition of the pineal organ of trout, and particularly the molecular species composition of PI, is more similar to the composition of the retina than that of the brain. Molecular species are abbreviated as follows: e.g., 16∶0/22∶6 PC is 1-palmitoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine.     

Fatty acids in femaleMacracanthorhynchus hirudinaceus (Acanthocephala)

Neutral lipids and phospholipids in the body wall, lemnisci and pseudocoel and neutral lipids of fluid found in the “tube” system of the lemnisci and the lacunar system of the body wall ofMacracanthorhynchus hirudinaceus (Acanthocephala) were determined by the technique of thin layer chromtography and gas chromatography (GC). Sixteen different fatty acids from nonpolar lipids were identified as follows: 8∶0, 10∶0, 11∶0, 12∶0, 13∶0, 14∶0, 14∶1, 16∶0, 18∶0, 18∶1, 18∶2 and/or 20∶0, 18∶3 and/or 20∶1, 20∶3, 22∶1, 24∶1 and 22∶6. In addition, there were three unidentified GC peaks corresponding to chain lengths greater than 20 carbons. Sixteen different fatty acids from phospholipids were identified in each of the three fractions analyzed. They were as follows: 10∶0, 11∶0, 12∶0, 13∶0, 14∶0, 14∶1, 16∶0, 16∶1, 18∶0, 18∶1, 18∶2 and/or 20∶0, 20∶2, 20∶3, 22∶1, 24∶1, and 22∶6. Four unidentified peaks were also observed. There was a significant difference in the percentage of lipid as well as the concentration of particular fatty acids when each fraction was compared. There was also an abundant supply of sterols and glycerides in each fraction.     

Lipid,sterol and fatty acid composition of antarctic krill (Euphausia superba Dana)

The lipid classes, fatty acids of total and individual lipids and sterols of Antarctic krill (Euphausia superba Dana) from two areas of the Antarctic Ocean were analyzed by thin layer chromatography (TLC), gas liquid chromatography (GLC) and gas liquid chromatography/mass spectrometry (GLC/MS). Basic differences in the lipid composition of krill from the Scotia Sea (caught in Dec. 1977) and krill from the Gerlache Strait (caught in Mar. 1981) were not observed. The main lipid classes found were: phosphatidylcholine (PC) (33–36%), phosphatidylethanolamine (PE) (5–6%), triacylglycerol (TG) (33–40%), free fatty acids (FFA) (8–16%) and sterols (1.4–1.7%). Wax esters and sterol esters were present only in traces. More than 50 fatty acids could be identified using GLC/MS, the major ones being 14∶0, 16∶0, 16∶1(n−7), 18∶1(n−9), 18∶1(n−7), 20∶5(n−3) and 22∶6(n−3). Phytanic acid was found in a concentration of 3% of total fatty acids. Short, medium-chain and hydroxy fatty acids (C≤10) were not detectable. The sterol fraction consisted of cholesterol, desmosterol and 22-dehydrocholesterol.     

Effects of dietary protein and cholesterol on phosphatidylcholine and phosphatidylethanolamine molecular species in mouse liver

The present study examined the effects of two atherogenic factors, animal protein and cholesterol, on the distribution of fatty acids and the molecular species of major liver phospholipids in mice. Weanling mice were fed a semisynthetic diet supplemented with either casein or soy protein (20%, w/w) in the presence or absence of 0.5% cholesterol for 4 wk. Results from mouse liver showed that animal protein and, more so, dietary cholesterol modified the fatty acid profiles of the phospholipids. Animal protein had no significant effect on the concentration of lipids, but it altered the relative distribution and fatty acid profiles of the phospholipids, phosphatidylcholine and phosphatidylethanolamine. Dietary cholesterol, on the other hand, significantly increased the concentration of liver lipids, but it did not alter the relative distribution of phosphatidylcholine and phosphatidylethanolamine. In cholesterol-fed mice, the proportions of molecular species containing 18∶2n−6 were increased, whereas those containing 20∶4n−6 were decreased, indicating that dietary cholesterol suppressed linoleic acid metabolism. Since cholesterol feeding selectively decreased the ratio of 18∶0/20∶4n−6 in phosphatidylcholine, whereas it increased the 18∶0/18∶2n−6 ratio in phosphatidylethanolamine, this finding suggests that dietary cholesterol may affect the incorporation of fatty acids but not the rate of synthesis of phosphatidylcholine and phosphatidylethanolamine.     

Long chain fatty acid deficits in brain myelin sphingolipids of undernourished rat pups

A restricted maternal dietary intake (40% of ad libitum intake) is known to cause myelin deficit that is accompanied by decreased amounts of individual phospholipids and sphingolipids in brain myelin of suckling rats. This communication reports the effects of the same nutritional stress on the fatty acid composition of brain myelin lipids. In myelin of 19-day-old normally fed rats, palmitate (16∶0), stearate (18∶0) and oleate (18∶1) accounted for 80–90% of all fatty acids in phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine. Maternal dietary restriction resulted in deficits of total fatty acid content, but did not affect the proportional distribution of individual fatty acids among phospholipids. By contrast, longer chain (22- and 24-carbon) fatty acids accounted for more than half the fatty acid content of myelin cerebroside and sulfatide from the 19-day-old control rat pups. In undernourished rats of that age, proportions of lignocerate (24∶0) and nervonate (24∶1) in cerebroside and sulfatide were 40–50% lower than those in control rats. This, together with higher proportions of 16∶0, 18∶0 and 18∶1 and a higher ratio of C₁₆−C₂₀ to C₂₂−C₂₄ in under-nourished than in control rats, suggests an impairment in fatty acid chain elongation. Ten days of nutritional rehabilitation failed to restore the fatty acid imbalances; however, after an additional 5 days of ad libitum feeding, the experimental and control values were similar. The undernutrition results in hypomyelination, which is characterized by a proportional decrease in lignoceric and nervonic acids of sphingolipids.     

Influence of partially hydrogenated vegetable and marine oils on lipid metabolism in rat liver and heart

Partially hydrogenated marine oils containing 18∶1-, 20∶1- and 22∶1-isomers and partially hydrogenated peanut oil containing 18∶1-isomers were fed as 24–28 wt % of the diet with or without supplement of linoleic acid. Reference groups were fed peanut, soybean, or rapeseed oils with low or high erucic acid content. Dietary monoene isomers reduced the conversion of linoleic acid into arachidonic acid and the deposition of the latter in liver and heart phosphatidylcholine. This effect was more pronounced for the partially hydrogenated marine oils than for the partially hydrogenated peanut oil. The content oftrans fatty acids in liver phospholipids was similar in groups fed partially hydrogenated fats. The distribution of various phospholipids in heart and liver was unaffected by the dietary fat. The decrease in deposition of arachidonic acid in rats fed partially hydrogenated marine oils was shown in vitro to be a consequence of lower Δ6-desaturase activity rather than an increase in the peroxisomal β-oxidation of arachidonic acid. The lower amounts of arachidonic acid deposited may be a result of competition in the Δ6-desaturation not only from the C22-and C20-monoenoic fatty acids originally present in the partially hydrogenated marine oil, but also from C18- and C16-monoenes produced by peroxisomal β-oxidation of the long-chain fatty acids. Part of this work was presented at the ISF-AOCS Congress, New York City, 1980.     

The fatty acid and aldehyde composition of the major phospholipids of mouse brain

Phospholipid classes were separated from mouse brain lipid extracts by preparative thin-layer chromatography (TLC). Methyl esters were prepared from the intact phospholipids by direct transesterification at room temperature in the presence of silica gel by using 0.5m NaOH-methanol in order to prevent interference by aldehydes or derivatives. Dimethyl acetal derivatives of phosphoglyceride alkenyl ethers (alkenyl moiety with a double bond in 1,2-position relative to oxygen linkage) were prepared, using 5% concentrated HCl in methanol, followed by preparative TLC for isolation. The major phospholipids present were ethanolamine phosphoglycerides (EPG) 39.8%, choline phosphoglycerides (CPG) 39.7%, serine phosphoglycerides (SPG) 15.0%, and sphingomyelin (Sph) 5.4%. One-fifth of the total phospholipids (PL) were in the form of plasmalogens, mainly EPG. Choline and serine plasmalogens were present in trace quantities. The major aldehyde components of the plasmalogens were 16∶0, 18∶0, and 18∶1. The EPG were rich in long-chain poly-unsaturated fatty acids, including 28.8% of 22∶6 and 17.0% of 20∶4, but contained only 7.2% of 16∶0. In contrast, the CPG contained 39.6% of 16∶0, and 31.0% of 18∶1 with a small content of polyunsaturated fatty acids. The SPG exhibited a still different pattern containing 38.2% of 18∶0, 23.2% of 18∶1, 24.3% of 22∶6, 2.9% of 16∶0, and 3.8% of 20∶4. Presented in part at the AOCS Meeting, New Orleans, May 1967.