Lactosylceramide: effect of acyl chain structure on phase behavior and molecular packing

Lactosylceramide (LacCer) is a pivotal intermediate in the metabolism of higher gangliosides, localizes to sphingolipid-sterol "rafts," and has been implicated in cellular signaling. To provide a fundamental characterization of LacCer phase behavior and intermolecular packing, LacCer containing different saturated (16:0, 18:0, 24:0) or monounsaturated (18:1(Delta9), 24:1(Delta15)) acyl chains were synthesized and studied by differential scanning calorimetry and Langmuir film balance approaches. Compared to related sphingoid- and glycerol-based lipids, LacCers containing saturated acyl chains display relatively high thermotropic and pressure-induced transitions. LacCer monolayer films are less elastic in an in-plane sense than sphingomyelin films, but are somewhat more elastic than galactosylceramide films. Together, these findings indicate that the disaccharide headgroup only marginally disrupts gel phase packing and orients more perpendicular than parallel to the interface. This contrasts the reported behavior of digalactosyldiglycerides with saturated acyl chains. Introducing single cis double bonds into the LacCer acyl chains dramatically lowers the high thermotropic and pressure-induced transitions. Greater reductions occur when cis double bonds are located near the middle of the acyl chains. The results are discussed in terms of how an extended disaccharide headgroup can enhance interactions among naturally abundant LacCers with saturated acyl chains.     

Body size related adaptations of the avian myocardial phospholipid fatty acyl chain composition

The myocardial phospholipid fatty acid (FA) composition (mol %) of 7 avian species was determined, in a body mass range from 150 g (Japanese quail, Coturnix coturnix japonica) to 19 kg (turkey, Meleagris gallopavo). Significant allometric increases were found for C16:1 n7 (allometric exponent: B = 0.15), C18:1 n7 (B = 0.08), C18:1 n9 (B = 0.24), C20:1 n9 (B = 0.22) and C20:3 n3 (B = 0.12); moreover, total monounsaturates (B = 0.20) and the sum of n9 FAs (B = 0.24) was also positively related to body mass. The total n3 FAs (B = − 0.36), and within them C22:5 n3 (B = − 0.41) and C22:6 n3 (B = − 0.60) showed allometric declines, such as total polyunsaturated fatty acids (PUFA; B = − 0.01), unsaturation index (B = − 0.03) and mean FA chain length (B = − 0.003). Comparing our results with earlier published data on avian skeletal muscle and divergent mammalian tissues, the allometric scaling of the above membrane forming fatty acids seems to be part of a general relationship postulated as the theory “membranes as metabolic pacemakers”. In addition, the cardiac muscle malondialdehyde concentration was negatively related to body mass (B = − 0.16), referring to a lower level of lipid peroxidation in larger birds, and vice versa, indicating a progressive myocardial lipid peroxidation in smaller-bodied species.     

Dependence of Escherichia coli hyperbaric oxygen toxicity on the lipid acyl chain composition.

This study examines certain membrane-related aspects of oxygen poisoning in Escherichia coli K1060 (fabB fadE lacI) and its parent strain, K-12 Ymel. Cells were grown to exponential or stationary phase in a minimal medium and exposed to air plus 300 lb/in2 of O2 as a suspension in minimal salts. After an initial lag, both strains lost viability with apparent first-order kinetics. Hypebaric oxygen was more toxic to cells harvested during the exponential phase of growth than to cells harvested from the stationary phase of growth for both strains K-12 Ymel and K1060. Control suspensions exposed to air plus 300 lb/in2 of N2 did not lose viability during a 96-h exposure. The sensitivity of the unsaturated fatty acid auxotroph, strain K1060, to hyperbaric oxygen increased as the degree of unsaturation of the fatty acid supplement increased. Cells grown with a cyclopropane fatty acid (9,10=methylenoctadecanoate) were the most resistant; cells grown with a monounsaturated fatty acid (oleate) were intermediate; and those grown with polyunsaturated fatty acids (linoleate and linolenate) were most sensitive to hyperbaric oxygen. The parent strain, K-12 Ymel, lost viability in hyperbaric oxygen most similarly to strain K1060 supplemented with oleate. To determine the relative effect of hyperbaric oxygen on the survival of E. coli with saturated membranes, substrains of K1060 were selected for growth on 12-methyltetrade-canoate or on 9 or 10-monobromostearate. Substrains grown with a saturated fatty acid supplement were equally or more sensitive to hyperbaric oxygen than when the same substrains were grown with a cyclopropane fatty acid supplement. The lipid acyl chain composition was determined in E. coli K1060 before and after exposure to hyperbaric oxygen or hyperbaric nitrogen. The proportion of nonsaturated acyl chain lipid of either the oleate- or the 9,10-methyleneoctade-canoate-supplemented K1060 remained unchanged after hyperbaric gas exposure. In strain K1060 supplemented with linoleate and grown to stationary phase, however, the relative unsaturated acyl chain content after hyperbaric exposure decreased in both gases. This finding prompted an investigation of the role of lipid oxidation in hyperbaric oxygen toxicity. Assays of potential lipid oxidation products were performed with linoleate-grown cells. The lipid hydroperoxide and peroxide content of the lipid extract increased by 6.9 times after 48 h of air plus 300 lb/in2 of O2; malondialdehyde and fluorescent complex lipid oxidation products showed much smaller or no changes. Lipid extracts from hyperbaric oxygen-exposed cells were not toxic to viable E. coli K1060, nor did they increase the rate of loss of viability in cells simultaneously exposed to hyperbaric oxygen. Linoleic acid hydroperoxide at 1.0 mM had no effect on the viability of E. coli K-12 Ymel and only marginally decreased the viability of E. coli K1060 supplemented with linoleate. We conclude that the kinetics of oxygen toxicity in E...     

Sphingomyelin modulates interfacial binding of Taiwan cobra phospholipase A2

The goal of the present study is to elucidate the effect of sphingomyelin on interfacial binding of Taiwan cobra phospholipase A₂ (PLA₂). Substitution of Asn-1 with Met caused a reduction in enzymatic activity and membrane-damaging activity of PLA₂ toward phospholipid vesicles, while sphingomyelin exerted an inhibitory effect on the biological activities of native and mutated PLA₂. Incorporation of sphingomyelin reduced membrane fluidity of phospholipid vesicles as evidenced by Laurdan fluorescence measurement. The results of self-quenching studies, binding of fluorescent probe, trinitrophenylation of Lys residues and fluorescence energy transfer between protein and lipid revealed that sphingomyelin altered differently membrane-bound mode of native and mutated PLA₂. Moreover, it was found that PLA₂ and N-terminally mutated PLA₂ adopted different conformation and geometrical arrangement on binding with membrane bilayer. Nevertheless, the binding affinity of PLA₂ and N-terminal mutant for phospholipid vesicles was not greatly affected by sphingomyelin. Together with the finding that mutation on N-terminus altered the gross conformation of PLA₂, our data indicate that sphingomyelin modulates the mode of membrane binding of PLA₂ at water/lipid interface, and suggest that the modulated effect of sphingomyelin depends on inherent structural elements of PLA₂.     

Alteration of the acyl chain composition of free fatty acids,acyl coenzyme A and other liPids by dietary polyunsaturated fats

Dietary alterations were used to demonstrate selective handling of fatty acids during their redistributionin vivo. Differences in the mol Per cent of individual acyl chains in the non-esterified fatty acid, acyl-coenzyme A and PhosPholiPid fractions reflected a result of relative Precursor abundance combined with enzymic selectivities. Selective distributions were observed in the utilization of individual acyl chains between 16:0 and 18:0, 18:1 and 18:2, and among 20:3, 20:4 and 20:5, 22:6 by ligase(s), hydrolase(s) and acyl-transferases. The variations in the mol Per cent of linoleate Present in the acyl-coenzyme A fraction of liver relative to that in the non-esterified fatty acids suggested anin vivo regulation of the level of linoleoyl-coenzyme A that influenced the synthesis of both arachidonoyl-coenzyme A and lipids. The greater abundance of eicosaPentaenoic acid in the free fatty acid fraction relative to that in the acyl-coenzyme A fraction may increase the ability of dietary 20: 5n-3 to be an effective inhibitor of the synthesis of Prostaglandins derived from 20:4n-6.     

Sphingomyelin analogs with branched N-acyl chains: The position of branching dramatically affects acyl chain order and sterol interactions in bilayer membranes

Sphingolipids have been found to have single methyl branchings both in their long-chain base and in their N-linked acyl chains. In this study we determined how methyl-branching in the N-linked acyl chain of sphingomyelin (SM) affected their membrane properties. SM analogs with a single methyl-branching at carbon 15 (of a 17:0 acyl chain; anteiso) had a lower gel-liquid transition temperature as compared to an iso-branched SM analog. Phytanoyl SM (methyls at carbons 3, 7, 11 and 15) as well as a SM analog with a methyl on carbon 10 in a hexadecanoyl chain failed to show a gel-liquid transition above 10 °C. Only the two distally branched SM analogs (iso and anteiso) formed ordered domains with cholesterol in a 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) bilayer. However, domains formed by the branched SM analogs appeared to contain less sterol when compared to palmitoyl SM (PSM) as the saturated phospholipid. Sterol-enriched domains formed by the anteiso SM analog were also less stable against temperature than domains formed by PSM. Both the 10-methyl and phytanoyl SM analogs failed to form sterol-enriched domains in the POPC bilayer. Acyl chain branching weakened SM/sterol interactions markedly when compared to PSM, as also evidenced from the decreased affinity of cholestatrienol to bilayers containing branched SM analogs. Our results show that methyl-branching weakened intermolecular interactions in a position-dependent manner.     

Survival of rabbit and horse erythrocytes in vivo after changing the fatty acyl composition of their phosphatidylcholine

The phospholipid composition and the distribution of phospholipids over the two leaflets of the membrane have been investigated for rabbit and horse erythrocyte membranes. Phosphatidylcholine (PC) comprises 39.4% and 41.3% of the total phospholipid complement of the rabbit and horse erythrocytes, respectively. In both membranes the distribution of this phospholipid is asymmetric: 70% of the PC is present in the outer layer of the rabbit membrane and 60% in that of the horse. The major species of this phospholipid class are the (1-palmitoyl-2-oleoyl)- and the (1-palmitoyl-2-linoleoyl)PC. The disaturated species, (1,2-dipalmitoyl)PC, is present in limited amounts only. Partial replacement of the native PC from intact erythrocytes was accomplished with a purified PC specific transfer protein from bovine liver. Replacement of the native PC species with (1-palmitoyl-2-oleoyl)PC up to 40% of the total PC complement had no effect on the osmotic fragility, the shape and the in vivo survival time of both erythrocyte species. Replacement of the native PC in both rabbit and horse erythrocytes with (1,2-dipalmitoyl)PC up to 20% gave rise to an increased osmotic fragility, a shape change from discocytic to echinocytic and a significant reduction in survival time measured after reinjection of the modified cells. At 30% replacement with (1,2-dipalmitoyl)PC the resulting spheroechinocytes appeared to be cleared from the circulation within 24 h after reinjection. The conclusion can be drawn that the repair mechanisms which may exist in vivo are insufficient to cope with the drastic changes in properties of the erythrocyte membrane which are induced by replacing more than 15% of the native PC by the dipalmitoyl species.     

Alanine transport by Chinese hamster ovary cells with altered phospholipid acyl chain composition

The Na+-dependent transport of alanine has been examined in Chinese hamster ovary (CHO) cells as a function of the fatty acid composition of their membrane lipids. Significant changes in the fatty acid composition of the CHO cell phospholipids were achieved by supplementation of the growth medium with specific saturated (palmitate) or monoenoic (oleate) free fatty acids. Arrhenius plots of the temperature-dependent uptake of alanine were constructed for cells of altered fatty acid composition. Alanine uptake was characterized by a single discontinuity in the Arrhenius plot. The temperature of this break was observed to be dependent upon the fatty acid composition of the cell phospholipids, ranging from 16 degrees C for cells enriched with oleate to 32 degrees C for cells enriched in palmitate. Calculation of the Km value for the uptake process showed no significant change with temperature or fatty acid supplementation. Correlations are made between the physical state of the membrane lipids and the temperature-dependence for alanine transport. The results are discussed in terms of membrane fatty acid composition, ordered in equilibrium fluid phase transitions and amino acid transport.     

Thermotropic phase behavior of mixed-chain phosphatidylglycerols: implications for acyl chain packing in fully hydrated bilayers

In this communication we report the first systematic investigation of the thermodynamic properties of fully hydrated mixed-chain phosphatidylglycerols (PG) using high-resolution differential scanning calorimetry (DSC). The crystal structure of dimyristoylphosphatidylglycerol shows an acyl chain conformation that is nearly opposite to that of phosphatidylcholine (PC). In PC, the sn-1 chain is straight while the sn-2 chain contains a bend; for PG, the sn-1 contains a bend while the sn-2 chain is in the all-trans conformation (R.H. Pearson, I. Pascher, The molecular structure of lecithin dihydrate, Nature, 281 (1978) 499-501; I. Pascher, S. Sundell, K. Harlos, H. Eibl, Conformational and packing properties of membrane lipids: the crystal structure of sodium dimyristoylphosphatidylglycerol, Biochim. Biophys. Acta, 896 (1987) 77-88). If the structure of PG found in the single crystal can be extrapolated to that in the fully hydrated gel-state bilayer, the observed difference in acyl chain conformations implies that modulation of the acyl chain asymmetry will have an opposite effect on the thermotropic phase behavior of PG and PC. For example, it is expected, based on the crystal structures, that C(15):C(13)PG should have a higher main phase transition temperature (Tm) than C(14):C(14)PG, and C(13):C(15)PG should have a lower Tm than C(14):C(14)PG. However, our DSC studies show clearly that the expectation is not borne out by experimental data. Rather, the Tm values of C(15):C(13)PG, C(14):C(14)PG, and C(13):C(15)PG are 18.2 degrees C, 23.1 degrees C, and 24.4 degrees C, respectively. Several other PGs, each with a unique acyl chain composition, have also been studied in this laboratory using high-resolution DSC. It is shown that the acyl chain conformation of fully hydrated PG in general is nearly opposite to that seen in the PG crystal structure.     

Metabolism of phosphatidylcholine and its implications for lipid acyl chain composition in Saccharomyces cerevisiae

Phosphatidylcholine (PC) is a very abundant membrane lipid in most eukaryotes including the model organism Saccharomyces cerevisiae. Consequently, the molecular species profile of PC, i.e. the ensemble of PC molecules with acyl chains differing in number of carbon atoms and double bonds, is important in determining the physical properties of eukaryotic membranes, and should be tightly regulated. In this review current insights in the contributions of biosynthesis, turnover, and remodeling by acyl chain exchange to the maintenance of PC homeostasis at the level of the molecular species in yeast are summarized. In addition, the phospholipid class-specific changes in membrane acyl chain composition induced by PC depletion are discussed, which identify PC as key player in a novel regulatory mechanism balancing the proportions of bilayer and non-bilayer lipids in yeast.     

Calcium alters the acyl chain composition and lipid fluidity of rat hepatocyte plasma membranes in vitro

Calcium ion decreases the lipid fluidity of isolated rat hepatocyte plasma membranes by modulating the activity of membrane enzymes which alter the lipid composition. To explore the mechanism of the effect of the cation, eight fluorophores were used to assess lipid fluidity via estimations of either steady-state fluorescence polarization or excimer fluorescence intensity. The results demonstrate that the reduction in fluidity occurs in the hydrophobic interior of the bilayer and that both the dynamic and static (lipid order) components of fluidity are affected by treatment with calcium. Analysis of the membrane lipids demonstrates that calcium treatment decreases the arachidonic acid content of the polar lipid fraction and, thereby, reduces the double-bond index of the fatty acids. This change in composition, which is expected to reduce the lipid fluidity, may result from activation by calcium of the endogenous hepatocyte plasma membrane phospholipase A2.     

Chlorpromazine associates with phosphatidylserines to cause an increase in the lipid's own interfacial molecular area--role of the fatty acyl composition

Partition coefficients of the drug chlorpromazine were determined for five different molecular species of diacylglycerophosphatidylserine in a monolayer kept at constant surface pressure (20 mN/m). Two models of adsorption of chlorpromazine in phosphatidylserine monolayers were compared. The first model correlated the amount of inserted drug molecules with the induced increase in area. The second model introduced the effect of drug adsorption on the lipid's own area by comparing the effect of increasing temperature on the lipid's own interfacial area. From the second model, the extrapolated work of insertion of one drug molecule per lipid molecule in a monolayer kept at 20 mN/m was correlated to the partition of the drug in liposomes. The work of insertion of chlorpromazine was insignificant in the unsaturated dioleoylphosphatidylserine and was maximum in the saturated distearoylphosphatidylserine monolayers. The presence of one double bond in the acyl chains dramatically reduces the work of insertion of chlorpromazine between lipid molecules and also reduces the effect chlorpromazine induces on the lipids own interfacial area in monolayers.     

Quantitative determination of acyl chain composition of subnanomole amounts of cellular long-chain acyl-coenzyme A esters

A procedure for the quantitative determination of the acyl chain composition of cellular long-chain acyl-CoA esters in subnanomole amounts is described. The abundant cellular lipids of samples are removed by extraction with organic solvents, and the proteins are precipitated from the aqueous phase by the addition of acetonitrile. The CoA thiolesters are adsorbed on neutral aluminum oxide and reduced with sodium borohydride to the corresponding alcohols that are then converted to t-butyldimethylsilyl ethers and analyzed quantitatively by gas chromatography. Saturated and unsaturated acyl chains behaved similarly throughout the procedure, and the common lipid esters do not interfere with the analysis of the CoA esters in the final assay procedure described. This simple and relatively rapid method is suitable for analyzing a large number of samples at a time.     

Influence of lecithin acyl chain composition on the kinetics of exchange between chylomicrons and high density lipoproteins

The kinetics of lecithin exchange between native lipoproteins was characterized for individual molecular species of lecithins of rat mesenteric lymph chylomicrons and rat plasma HDL. Studies were performed in the absence of lipid transfer proteins. Donor (chylomicrons) and acceptor (HDL) particles were present in ratios of 1:1 and 1:10 with respect to total phospholipid. Biphasic exchange kinetics were observed for all major lecithins common to chylomicrons and HDL at both proportions of donor to acceptor particles. During the early rapid phase of exchange, complete in about 30 min, 40-60% of the total lecithin pool was exchanged. Initial exchange rates were most rapid for the more hydrophilic species of the major lecithins normally present in both lipoproteins. Calculated activation energies correspondingly were least for a diunsaturated lecithin (18:1-20:4), intermediate for lecithins were 16:0 in position-1 (16:0-18:2 and 16:0-20:4), and highest for analogous lecithins with 18:0 in position-1. A 10-fold increase in the ratio of acceptor to donor particles affected neither the biphasic nature of the exchange nor the rates of exchange of individual molecular species (consistent with exchange by diffusion rather than by particle collisions). Total equilibration of individual molecular lecithin species was achieved by 24 hr (37 degrees C, donor to acceptor ratio of 1:1) with only a small change in the relative mass of lecithins in chylomicrons and HDL. Novel lecithins containing 18:3, incorporated into chylomicrons, were found to exchange exceedingly rapidly.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of acyl chain composition on salt-induced lamellar to inverted hexagonal phase transitions in cardiolipin

Salt-induced fluid lamellar (L alpha) to inverted hexagonal (HII) phase transitions have been studied in diphosphatidylglycerols (cardiolipins) with different acyl chain compositions, using 31P nuclear magnetic resonance (NMR) spectroscopy. Cardiolipins with four myristoyl chains, tetramyristoyl cardiolipin (TMCL), and with four oleoyl chains, tetraoleoyl cardiolipin (TOCL), were synthesized chemically. TMCL was found to undergo a thermotropic lamellar gel to lamellar liquid-crystalline phase transition at 33-35 degrees C. This lipid exhibited an axially symmetric 31P-NMR spectrum corresponding to a lamellar phase at all NaCl concentrations between 0 and 6 M. In the case of TOCL, formation of an HII phase was induced by salt concentrations of 3.5 M NaCl or greater. These observations, taken together with earlier findings that bovine heart cardiolipin aqueous dispersions adopt an HII phase at salt concentrations of 1.5 M NaCl or greater, indicate that increasing unsaturation and length of the acyl chains favour formation of the HII phase in diphosphatidylglycerols.     

Synthesis and interfacial behavior of three homologous glycero neoglycolipids with various chain lengths

Three neoglycolipids 1a–c derived from glycerol were synthezised and their molecular arrangements were studied at the air/water interface through Langmuir–Blodgett technique. The common structural features of these neoglycolipids are: a triethyleneglycol spacer at C-2 of glycerol, a GlcNAc hydrophilic head group at the end of the spacer, two saturated aliphatic chains at C-1 and C-3 of glycerol, linked by ether bonds. Compounds 1a–c differ only by the length of their lipid moieties. By increasing the hydrocarbon chain length from C₁₁ to C₁₆, a densely packed state was reached in the monolayer. The compression isotherms displayed an expanded state during the whole compression for compounds 1a and 1c bearing two C₁₁ or one C₁₁ and one C₁₆ chains. Compound 1b bearing two C₁₆ chains displayed a quite different interfacial behavior. The transfer of these monolayers onto a solid substrate can be obtained only with a trigger molecule, a phosphatidic acid.     

Crystallization of monoacylated proteins: influence of acyl chain length

The crystallization of monoacylated proteins has been investigated using a model system. Acylated derivatives of bovine pancreatic ribonuclease A, differing in their acyl chain lengths (10 to 16 carbon atoms), have been prepared using reverse micelles as microreactors. With one fatty acid moiety per polypeptide chain, covalently attached to the NH₂ terminus of the protein, all the modified proteins have similar enzymatic activity and hydrodynamic radius as the native protein. Only the caprylated derivative can give crystals which diffract to high resolution. The resolved structure indicates that: (i) the protein folding is not modified by the chemical modification, (ii) the capryl moiety is not buried within the molecule but available for external interactions. Dynamic light scattering experiments on concentrated solutions show that protein-protein interactions are dependent on acyl chain length. Proteins with the longest attached chains (14 and 16 carbon atoms) tend to self-associate through acyl group interactions. Received: 4 October 1996 / Accepted: 13 December 1996     

Glycosphingolipids: 2H NMR study of the influence of carbohydrate headgroup structure on ceramide acyl chain behavior in glycolipid-phospholipid bilayers

Galactosyl- and glucosylceramide, globoside, and dihydrolactosylceramide, bearing [2,2-2H2]stearic acid, have been studied at a concentration of 10 mol% in bilayers of dimyristoylphosphatidylcholine by 2H NMR. The quadrupolar splitting delta vQ of the C2 deuterons were measured at several temperatures in the range of 30-60 degrees C. Spin-lattice relaxation times T1 of C2 deuterons were determined in the same temperature range for all lipids but globoside. T1 values at 30 and 50 degrees C were unexpectedly short (6-8 ms), indicating reduced mobility of the ceramide acyl chains compared to that of the host phospholipid. At all temperatures, both delta vQ and T1 were essentially identical for the monoglycosylated species, GalCer and GlcCer, indicating that the order and dynamics of the upper portion of the fatty acyl chain are insensitive to this small change in the headgroup structure. In the case of globoside, where the glycolipid headgroup is equivalent to that of GlcCer extended by three sugar residues, values for the quadrupolar splittings associated with the acyl chain C2-position were very close to those obtained for Gal- and GlcCer. In contrast, the delta vQ values obtained for the diglycosyl species, LacCer, were significantly different at all temperatures. This different behavior of LacCer relative to that of the other glycolipids most likely originates from an orientational change of the acyl chain at the C2-position due to the absence of a 4,5 double bond in dihydrosphingosine. T1 values for the GlcCer and GalCer systems increased with temperature, indicating that the motions responsible for relaxation were in the short correlation time regime.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ontogenic development of the fatty acyl chain composition of the turkey (Meleagris gallopavo) pectoralis superficialis muscle membranes: an allometric approach

The growth-associated development of the m. pectoralis superficialis (MPS) phospholipid (PL) and triacylglycerol (TAG) fatty acyl (FA) chain composition was determined in BUT8 meat-type turkeys. Samples (3 d, 8, 12, 16 and 20 wk) of each 6 males were analysed by lipid fractionation and subsequent gas chromatography. Results were interpreted on an allometric basis. The MPS mass increased linearly (MPS weight = 0.2787 BW- 123.67; R2 = 0.9935, P<0.001, n = 30). In the total phospholipids 62-63% unsaturated fatty acids were found irrespective of the diet. A negative allometric alteration was found for the total saturated acyl chains (B = -0.012), while a positive value for the calculated unsaturation index (B = 0.026) was obtained. Within the PUFA chains, the n3- n6 balance was markedly changed, on the favour of the n3 fatty acyl chains, namely competitive allometric trends were found for the total n3 (B = 0.087) and n6 (B = 0.032) fatty acid groups. The alterations of the TAG FA chain composition were diet-dependent. The serum creatine kinase activity increased by over one class of magnitude during the trial. The allometric approach was found to be powerful in the characterization of the basic, non diet-dependent ontogenic alterations of the phospholipid fatty acyl chain composition.