Lipid class composition of the protozoan Perkinsus marinus, an oyster parasite, and its metabolism of a fluorescent phosphatidylcholine analog

Perkinsus marinus is one of two important protozoan parasites of the eastern oyster, Crassostrea virginica. The other is Haplosporidium nelsoni. Lipids extracted from 7-d-old in vitro cultured P. marinus meronts, incubated with fluorescent-labeled phosphatidylcholine (FL PC) and nonincubated P. marinus meronts, were analyzed by a high-performance liquid chromatography (HPLC) system equipped with a diol phase column, in combination with thin-layer chromatography coupled with a flame-ionization detector (TLC/FID), and high-performance thin-layer chromatography (HPTLC). Various polar and neutral lipid classes were separated by HPLC using a two-gradient solvent system. Five polar lipid classes--phosphatidylcholine (PC), phosphatidylethanolamine (PE), cardiolipin (CL), sphingomyelin (SM), and phosphatidylserine (PS)--were identified from P. marinus extracts. Four neutral lipid classes--triacylglycerol (TAG), steryl ester (SE), cholesterol (CHO), and fatty alcohol--were distinguished. TLC/FID analysis of meront lipids showed that the weight percentages of PC, PE, CL, SM, PS/PI, TAG, SE, and CHO were 21, 10.7, 4, 2.3, 4.3, 48.7, 7.8, and 1.2%, respectively. HPLC and HPTLC analyses revealed the presence of two SM and PS isomers in P. marinus extracts. Perkinsus marinus effectively incorporated FL PC acquired from the medium and metabolized it to various components (i.e., free fatty acid, monoacylglycerol, diacylglycerol, TAG, PE, and CL). Uptake and interconversion of FL PC in P. marinus meronts increased with time. After 48 h the total uptake of fluorescence (FL) was 28.9% of the FL PC added to the medium, and 43% of the incorporated FL resided in TAG.     

Lipid class and fatty acid composition of the boreal soft coral Gersemia rubiformis

Total lipid, phospholipid, and FA composition and distribution of FA between polar lipids (PL) and neutral lipids (NL) were investigated in the boreal soft coral Gersemia rubiformis from the Bering Sea. The total lipids were mostly hydrocarbons and waxes (33.7%) and PL (33.1%). The content of monoalkyldiacylglycerols (9.7%) exceeded the content of TAG (6.7%). PC and PE constituted 31.4% and 25.6% of total phospholipids, respectively. Principal FA were 16∶0, 16∶1n−7, 18∶0, 18∶1n−9, 18∶1n−7, 20∶1n−7, 20∶4n−6, 20∶4n−3, 20∶5n−3 22∶5n−3, 22∶6n−3, 24∶5n−6, and 24∶6n−3. Most n−6 PUFA (52% of total FA) were associated with the PL fraction; this was especially true for arachidonic and tetracosapentaenoic acids. The NL were enriched with mono-, di-, trienoic, and n−3 PUFA. The variation in EPA levels in both NL and PL suggests an origin of this acid from lipids of diatoms consumed by the corals.     

Lipid class and fatty acid composition of intact peripheral nerve and during wallerian degeneration

Lipid extracts from normal cat, chicken, and beef sciatic nerve were fractionated into their components by combinations of silicic acid, Florisil, DEAE-cellulose, or silicic acid-silicate column chromatography. The constitutent fatty acids of total lipid extracts and of individual lipid classes were qualitatively and quantitatively determined as their methyl esters by gas chromatography. These methods were also applied to lipid extracts from cat sciatic nerve undergoing Wallerian degeneration at 8, 16, 32, and 96 days after section and to chicken sciatic nerve undergoing demyelination due to organophosphate poisoning. All fatty acids were markedly decreased in the total lipids of cat sciatic nerve at 96 days after section and most of these were decreased at 32 days. As early as 8 days after section 16:0, 16:1, 18:2, 20:0, and 20:4 showed decreases, while 18:0, 18:1, 22:1, 22:5, 22:6, and 24:1 did not begin to show decreases until 16 days after section. The decreases in fatty acids were considered to be due to increased catabolism, decreased synthesis, or increased removal of fatty acids from nervous tissue. The fatty acid content of the total lipids of chicken nerve undergoing demyelination resembled that of cat sciatic nerve between 16 and 32 days after section. Myelin lipids, sphingomyelin, cerebrosides, and phosphatidyl ethanolamine (PE) began to decrease as early as 8 days after section in cat sciatic nerve. Phosphatidyl serine (PS) also decreased at this time. Cholesterol, lecithin, and ethanolamine plasmalogen did not begin to decrease until 16 days after section and phosphatidyl inositol (PI) did not decrease until 32 days after section. Triglycerides decreased markedly at 8 days after section gradually returning to normal by 96 days. This was accompanied by a transient increase in free fatty acids and monoglycerides. Cholesterol esters and lysolecithin increased markedly at 8 days after section and were higher than normal levels even at 96 days after section. In chicken sciatic nerve undergoing demyelination after organophosphate poisoning, cerebroside was the only myelin lipid which decreased in amt, while cholesterol esters and diglycerides increased. Sphingomyelin and cerebrosides containing 16:0, 18:0, 18:1, 18:2, 20:0, 22:0, 23:0, 24:0, 24:1 seemed to be most susceptible to degradation or interference in synthesis in degenerating nerve. For the most part, these fatty acids were observed to increase in cholesterol esters, free fatty acids, and, in some instances, triglycerides. The changes in various lipid classes and their constituent fatty acids are discussed in relation to various cellular changes which accompany degeneration.     

Lipid composition of erythrocytes and thrombocytes of a subantarctic seabird, the king penguin

Phospholipid (PL) compositions and fatty acid (FA) patterns of PL were determined in the erythrocytes and blood thrombocytes of a seabird, the king penguin, living in the subantarctic area and feeding on prey rich in n−3 polyunsaturated FA. Results were compared between birds in three different physiological states (breeding and molting adults, chicks) to those reported for other birds. In erythrocytes, the ratios of cholesterol to PL and of sphingomyelin to phosphatidylcholine (PC) were lower than in other birds. The PL distribution was similar to those previously reported in the hen and pigeon. In contrast to other birds, cardiolipin levels were unexpectedly high (4%). Very long chain n−3 FA were abundant (13–27%) in phosphatidylethanolamine (PE), phosphatidylserine and PC, probably in relation to the natural diet of these birds. Among n−3 FA, 22∶6n−3 was the most abundant in all PL (2−20%), whereas the highest levels of arachidonic acid were observed in PE (14%). In thrombocytes, the PL distribution and FA composition of the main PL (PC, PE) differed from those of erythrocytes, and in particular, levels of n−3 FA (9–12%) were 1.5–2 times lower. The highest levels of arachidonic acid were found in phosphatidylinositol (24%). The lipid profile of penguin erythrocytes could contribute to the efficiency of blood circulation and oxygen delivery in microvascular beds, thus favoring diving capacity of these animals. Our observations do not support the hypothesis of a common origin of avian thrombocytes and erythrocytes.     

Lipid class composition during embryonic and early larval development in Atlantic herring (Clupea harengus,L.)

The lipid class compositions of Atlantic herring eggs and larvae were determined immediately before fertilization, after fertilization and at various times during subsequent embryonic and early larval development. Total lipid constituted 15% of the dry wt of ripe eggs, 70% of the total lipid being polar lipid with phosphatidylcholine (PC) accounting for almost 90% of the polar lipid. In general, the total lipid content decreased gradually during embryogenesis and in particular during larval development. Within 3 hr after fertilization the relative percentage of neutral lipid decreased slightly. This was followed by a general decrease in polar lipid which, by the stage of yolk sac absorption, was reduced to 52% of the total lipid. The decreased percentage of polar lipid was due entirely to a decrease in PC, which was reduced to 66% of the polar lipids at the stage of yolk sac absorption. The accompanying increase in the percentage of neutral lipids was mainly due to increased percentages of triacylglycerols (TAG) up to yolk sac absorption and cholesterol esters in the larval stages. During the first 4 days after hatching, phospholipids and to a lesser extent cholesterol were preferentially depleted in the yolk sacs, which also had higher levels of free fatty acids. The results are discussed in relation to possible roles of different lipids during embryonic and early larval development.     

Skin lipids. II. Lipid class composition of samples from various species and anatomical sites

The literature is reviewed and new data are presented for the lipid class analysis of samples of lipid from the following skin tissues and appendages: Total epidermis, stratum corneum, and living epidermis from the human leg, total epidermis (human sole), total epidermis (rat body), sebaceous glands (human scalp), Meibomian glands (human eyelid), preputial glands (rat), preen gland (domestic goose), scalp skin surface (human),vernix caseosa (human fetal body), and rat skin surface from the back. Lipid yields are also given for most of these samples. The results show that the composition of the sebaceous type excretion varies not only from species to species but for different anatomical sites within a given species. Noteworthy is the striking number of unidentified components. Evidence is presented for the existence of a wax diester in the sebaceous gland excretion of the rat.     

Lipid metabolism of the yellow clam,Mesodesma mactroides: 2-Polyunsaturated fatty acid metabolism

The fate of labeled linoleic, α-linolenic, and higher homologs of α-linolenic acid administered to the yellow clam,Mesodesma mactroides, was investigated. It was found that the clam incorporated the acids dissolved in sea water and converted 18∶2 (n−6) into 20∶2 (n−6) and 18∶3 (n−3) into 18∶4 (n−3) and 20∶3 (n−3). The addition of casein hydrolysate to the sea water increased the desaturation capacity of the clam and allowed the conversion of 18∶2 (n−6) into 18∶3 (n−6) to be demonstrated. An enhanced desaturation of 18∶3 (n−3) into 18∶4 (n−3) was also demonstrated. After 12 hr administration of the acid, no radioactivity was found in arachidonic, 20∶5 (n−3), or 22∶6 (n−3). Feeding the clams a culture ofPhaeodactylum tricornutum previously incubated with 1-¹⁴C-α-linolenic acid demonstrated that all the homologs of the α-linolenic series were found in the clam without any important changes. Six hour administration of labeled linolenic acid resulted in the incorporation of the acid into diglycerides and phospholipids. Member of the carrera del Investigador Cientifico of the Consejo Nacional de Investigaciones Cientificas y Tecnicas     

Lipid metabolism of the yellow clam,Mesodesma mactroides: 3-Saturated fatty acids and acetate metabolism

The fate of labeled palmitate, stearate, and acetate administered to the yellow clam,Mesodesma mactroides, was investigated. 1-¹⁴C palmitic and 1-¹⁴C stearic acids were oxidized to CO₂ to a limited extent. They were mainly incorporated in diacylglycerols and triacylglycerols and were converted to higher homologs. After administration, palmitic acid was converted to stearic and oleic acids, whereas administered stearic acid was converted to 18∶1, 18∶2, 20∶1, and 20∶2 acids. Labeled acetate was readily included by the clam in 12∶0, 14∶0, 14∶1, 15∶0, 16∶1, 16∶1, 16∶2, 18∶2, 18∶1, 18∶2, 20∶1, 20∶2, and 20∶3 acids.     

Lipid metabolism and FA composition in tissues of Eurasian perch Perca fluviatilis as influenced by dietary fats

Eurasian perch, Perca fluviatilis, were fed a semipurified fat-free diet for 4 wk, followed by a 16% feeding supplementation of either olive oil (OO), safflower oil (SO), linseed oil (LO), or cod liver oil (CLO) as the only lipid source in each diet for 10 wk. Significant reductions in total lipid of tissues were observed (31.4% in viscera, 66.7% in muscle, and 74.1% in liver) after feeding the fat-free diet. The SO-, LO-, and CLO-fed fish significantly increased lipid deposition in liver and viscera compared to fish fed the OO diet; however, muscle lipid levels were not significantly affected. Large amounts of dietary 18∶1n−9 were incorporated directly into tissue lipids when fish were fed the OO diet. The LO diet significantly elevated 18∶4n−3, 20∶5n−3, 22∶5n−3, and 22∶6n−3 in the liver compared to fish fed OO or SO diets, and the n−3/n−6 ratio was 16 times that of the SO group, with significantly high desaturation and elongation products of 18∶3n−3. These results suggest that Δ6 and Δ5 desaturases are highly active in Eurasian perch, and that the enzymes at this dietary n−3/n−6 ratio favor 18∶3n−3 over 18∶2n−6 as substrate. The SO diet significantly increased 18∶3n−6, 20∶3n−6, and 22∶5n−6 in the liver and significantly decreased EPA and DHA. This indicates that desaturation enzymes were not specifically favoring n−3 over n−6 acids in perch lipid metabolism, and that these elongation and desaturation enzymes were influenced by n−3 and n−6 FA content in the diet. The present study indicates that high tissue content of DHA in the muscle of Eurasian perch was attributable to the greater ability for n−3 acid bioconversion.     

Lipid class composition of normal human brain and variations in metachromatic leucodystrophy,Tay-Sachs,Niemann-Pick,chronic gaucher’s and Alzheimer’s diseases

Procedures suitable for obtaining representative samples of whole brain and of total grey and white matter of brain are presented and discussed. A procedure is described for the quantitative determination of lipid class distribution of human brain specimens utilizing in sequence : a cellulose column to separate gangliosides and nonlipid material from the remaining lipids, diethylaminoethyl (DEAE) cellulose column chromatography to separate the lipid classes into manageable groups, and finally quantitation of the lipid classes by thin-layer chromatography (TLC). TLC is made quantitative by correlating the amt of charring of spots on chromatograms with the amt of lipid present by means of transmission densitometry. The use of two-dimensional TLC for the analysis of brain lipids and its application to the study of pathological brain specimens is also described. The application of these procedures to the study of metachromatic leucodystrophy, Tay-Sachs, Niemann-Pick, and Alzheimer’s diseases and senile cerebral cortical atrophy is described and data are presented. In two cases of Alzheimer’s disease, a large reduction in fresh weight and total lipid of brain were found; the lipid class distribution of whole brain in one case and of total grey and total white matter in another were essentially normal. The lipid class distributions of the brain in metachromatic leucodystrophy, Tay-Sachs disease, and Niemann-Pick disease were shown to be similar to that of normal infant brain except that one sphingolipid was greatly increased in each disease (sulfatide in metachromatic leucodystrophy, one ganglioside in Tay-Sachs disease, and sphingomyelin in Niemann-Pick disease).     

Lipid composition of the pecan weevil,Curculio caryae (Horn)

The lipids of larvae, male adults, and female adults of the pecan weevil,Curculio caryae (Horn), were studied, and special attention was given the fatty acid composition. The larvae contained an unusually high amount of lipid material (40.6%), most of it concentrated in the neutral lipid fraction. Male and famale adults contained more conventional quantities, 5.8 and 8.2%, respectively. Oleic acid was the major fatty acid in the total and neutral lipids of all stages; linoleic acid was the most abundant in the phospholipid fractions.     

Lipid and fatty acid composition of turkey liver,skin and depot tissue

Total lipid and phospholipid contents of liver, skin and depot fat from yearling hen turkeys have been studied. Liver lipid averaged 88.5 mg/g wet tissue; skin, 385.0 and depot fat, 753.5. Phospholipids comprised 32.05% of total lipid of liver, but only 0.81% of skin and 0.46% of depot fat. Fatty acids of liver differed from those of skin or depot fat by larger amounts of 16∶0, 18∶0, 20∶4, 22∶0 and 24∶0, and smaller amounts of 16∶1, 18∶1, 18∶2, 18∶3 and 20∶0. Similarity existed between skin and depot fat. Journal Paper No. J-6473 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project No. 1696.     

Lipid composition of the membrane released after an in vitro acrosome reaction of epididymal boar sperm

Prior to fertilization, mammalian sperm must undergo the acrosome reaction, which involves modifications of the plasma and outer acrosomal membranes followed by vesiculation and release of the membranes. The membrane fraction that was released from caudal boar sperm undergoing an in vitro acrosome-like reaction was isolated and characterized with respect to density, marker enzymes and lipid composition. This membrane had a lower phospholipid/protein ratio (mg/mg) than the sperm plasma membrane, whereas both membranes had similar molar sterol/phospholipid ratios. The major phospholipid was sphingomyelin, followed by phosphatidylethanolamine and phosphatidylcholine, whereas in the plasma membrane the order was reversed; the two major phosphoglycerides contained alkylacyl and alkenylacyl species in addition to the diacyl species. The released membrane also contained lower amounts of cholesterol sulfate and unsaturated fatty acids than the plasma membranes. These results, in combination with our studies on the changes of the sperm membranes during maturation and acrosome reaction, will allow a better understanding of the mechanism of the sperm acrosome reaction.     

Lipid and FA composition of the pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis>: Influence of season and maturation

The lipid and FA composition of the total lipids of the pearl oyster Pinctada fucata martensii, in different seasons and in different areas, were analyzed to clarify its lipid physiology and to estimate the possible influence of its prey phytoplankton. TAG and sterols were the major components in the neutral lipids in all conditions, whereas high levels of phospholipids (PE and PC) were found in the polar lipids. The major FA in the TAG in all samples were 14∶0, 16∶0, and 18∶0 as saturated FA (saturates); 16∶1n−7, 18∶1n−9, and 18∶1n−7 as monoenoic FA (monoenes); and 20∶4n−6 (arachidonic acid: AA), 20∶5n−3 (EPA), and 22∶6n−3 (DHA) as PUFA. The major components found in the polar lipids were 16∶0 and 18∶0 as saturates; 22∶2n−9, 15 and 22∶2n−7, 15 as non-methylene-interrupted dienes (NMID), and AA, 22∶3n−6, 9, 15, EPA, and DHA as PUFA. Although it is a marine animal, characteristically high levels of AA were found in both the TAG and phospholipids. This result suggests that lipids of P. fucata may be influenced by those of its phytoplanktonic prey. The increase in levels of NMID from TAG to PE with a decrease in those of monoenes suggests that the tissues of this species are able to biosynthesize only the less unsaturated PUFA, such as NMID. In particular, NMID derivatives are considered to be biosynthesized in the PE; thus, they might play a particular role in the membrane, because NMID were characteristically localized only in the PE.     

Environmental temperature and metabolism of the molecular species of phosphatidylcholine in the tissues of the rainbow trout

The phospholipid composition and the fatty acid pattern of phosphatidylcholine from major tissues of the rainbow trout acclimated to 11° and 21°C were analyzed. A significant difference in the level of 20∶5ω3 fatty acid between gill, kidney, intestine and liver phosphatidylcholine is observed in both groups of animals. These differences correlate with a modification of the molecular species distribution in the phosphatidylcholine in these organs. In vivo incorporation of³²P into phospholipids of gills, kidney, intestine, liver and muscle was determined in trout acclimated to 11° and 21°C. Temperature acclimation specifically modify the relative specific radioactivity of the monoene and hexaene molecular species of phosphatidylcholine in gills, intestine, kidney and liver. It can be concluded from these results, that the metabolism of the diacylglycerol moiety and the coupled fatty acids of phosphatidylcholine differs with the tissue and the temperature. It is suggested that different associated mechanisms specific for each organ determine the fatty acid composition of phophatidylcholine required for its physiological functions at each temperature.     

Lipid metabolism of the yellow clam,Mesodesma mactroides: I. Composition of the lipids

The lipid composition of the yellow clam,Mesodesma mactroides, that lives in the northern beaches of the Buenos Aires province of Argentina was studied. The main nonpolar lipids are triglycerides and alkoxyglycerides. Phosphatidyl choline, phosphatidyl ethanolamine, and phosphatidyl serine are the main phospholipids. The predominant fatty acids are 16∶0, 16∶1ω7, 18∶0, 18∶1ω9, 20∶5ω3, and 22∶6ω3. The are mainly provided by the clam's food and stored in the hepatopancreas. The content of polyunsaturated acids increases in summer together with an increase in nonpolar lipids and is correlative with an increase in phytoplankton in the sea water. Sexual maturity modifies the lipid composition of gametes.     

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.     

Effect of dexamethasone on the fatty acid composition of total liver microsomal lipids and phosphatidylcholine molecular species

Dexamethasone depresses Δ6 and Δ5 and increases Δ9 desaturase and synthase activities. Therefore, we investigated the effect on the fatty acid composition of microsomal liver lipids and phosphatidylcholine (PtdCho) molecular species. After 15 d of treatment we found a notable decrease in arachidonic acid, a small decrease in stearic acid, and increases of linoleic, oleic, palmitoleic, and palmitic acids in liver microsomal total lipids and PtdCho. The study of the distribution of the PtdCho molecular species indicated that 18∶0/20∶4n−6, 16∶0/20∶4n−6, and 16∶0/18∶2n−6 predominated in the control animals. Dexamethasone, as expected because of its depressing effect on arachidonic acid synthesis and activation of oleic and palmitic acid synthesis, evoked a very significant decrease in 18∶0/20∶4n−6 PtdCho (P<0.001) and an important increase in 16∶0/18∶2n−6. The invariability of 16∶0/20∶4n−6 PtdCho could be related to the antagonistic effect of arachidonic and palmitic acid synthesis. PtdCho species containing oleic acid were not significant. The bulk fluidity and dynamic properties of the microsomal lipid bilayer measured by fluorometry using the probes 1,6-diphenyl-1,3,5-hexatriene and 4-trimethylammonium-phenyl-6-phenyl-1,3,5-hexatriene showed no significant modification, probably owing to a compensatory effect of the different molecular species, but changes of particular domains not detected by this technique are possible. However, the extremely sensitive Laurdan detected increased lipid packing in the less-fluid domains of the polar-nonpolar interphase of the bilayer, possibly evoked by the change of molecular species and cholesterol/phospholipid ratio. The most important effect found is the decrease of arachidonic acid pools in liver phospholipids as one of the corresponding causes of dexamethasone-dependent pharmacological effects.     

Lipid metabolism in plasma,liver, and adipose tissue of rats with experimental chronic nephrotic syndrome

Plasma, liver, and adipose tissue lipid composition and synthesis from [1-¹⁴C] acetate were studied three months following induction of nephrotic syndrome in rats by injection of antiglomerular basement membrane protein. Plasma triglyceride concentrations and specific radioactivities were elevated, and the triglycerides contained increased proportions of oleic acid. Plasma cholesterol and phospholipid concentrations were also increased, but free fatty acid levels were not. Liver triglyceride concentrations were decreased and incorporation of [1-¹⁴] acetate into liver triglycerides was also depressed below that of normal controls. Nephrotic rat liver triglycerides contained a higher proportion of oleic acid and lower arachidonic acid than did controls. Incorporation of [1-¹⁴C] acetate into adipose tissue lipids of the nephrotic rats was increased, and the proportion of palmitic acid was decreased. In the chronic nephrotic rat, the major source of the increased plasma triglycerides may be fatty acids mobilized from adipose tissue stores.