Maternal exposure to fish oil primes offspring to harbor intestinal pathobionts associated with altered immune cell balance.

Our previous studies revealed that offspring from rat dams fed fish oil (at 8% and 18% energy), developed impaired intestinal barriers sensitizing the colon to exacerbated injury later in life. To discern the mechanism, we hypothesized that in utero exposure to fish oil, rich in n-3 polyunsaturated fatty acid (PUFA), caused abnormal intestinal reparative responses to mucosal injury through differences in intestinal microbiota and the presence of naïve immune cells. To identify such mechanisms, gut microbes and naïve immune cells were compared between rat pups born to dams fed either n-6 PUFA, n-3 PUFA or breeder chow. Maternal exposure to either of the PUFA rich diets altered the development of the intestinal microbiota with an overall reduction in microbial density. Using qPCR, we found that each type of PUFA differentially altered the major gut phyla; fish oil increased Bacteroidetes and safflower oil increased Firmicutes. Both PUFA diets reduced microbes known to dominate the infant gut like Enterobacteriaceae and Bifidobacteria spp. when compared to the chow group. Uniquely, maternal fish oil diets resulted in offspring showing blooms of opportunistic pathogens like Bilophila wadsworthia, Enterococcus faecium and Bacteroides fragilis in their gut microbiota. As well, fish oil groups showed a reduction in colonic CD8+ T cells, CD4+ Foxp3+ T cells and arginase+ M2 macrophages. In conclusion, fish oil supplementation in pharmacological excess, at 18% by energy as shown in this study, provides an example where excess dosing in utero can prime offspring to harbor intestinal pathobionts and alter immune cell homeostasis.

Long chain omega-3 fatty acids: micronutrients in disguise.

Considerable information has accumulated to show that DHA and EPA have unique roles that differ from other n-3 fatty acids and the n-6 fatty acids, with increasing understanding of the mechanisms through which these fatty acids reduce risk of disease. DHA and EPA regulate hepatic lipid and glucose metabolism, but are present in foods of animal origin, which are generally high in protein with variable triglycerides and low carbohydrate. Biological activity at intakes too low to provide significant amounts of energy is consistent with the definition of a vitamin for which needs are modified by life-stage, diet and genetic variables, and disease. Recent studies reveal that DHA may play a central role in co-coordinating complex networks that integrate hepatic glucose, fatty acid and amino acid metabolism for the purpose of efficient utilization of dietary protein, particularly during early development when the milk diet provides large amounts of energy from fat.

Dietary long chain n-3 fatty acids are more closely associated with protein than energy intakes from fat.

The n-3 fatty acids, eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) regulate hepatic lipid and glucose metabolism; however, EPA and DHA are naturally present in human diets in foods of animal origin, which are generally high in protein with variable triglycerides and uniformly low amounts of carbohydrate. We used dietary information for 611 individuals of 1.5-66 years to address whether EPA and DHA are associated with protein, but not fat intake. EPA, DHA and arachidonic acid (20:4n-6) intakes were positively associated with protein, but not fat intake, whereas linoleic acid (18:2n-6) and α-linolenic acid (18:3n-3) intakes were positively associated with fat, but not protein intake. Children 1-3 years of age have lower EPA and DHA intakes than children over 4 years or adults. Recommendations regarding EPA and DHA intake should focus on protein sources, rather than diet fat, and consider their potential roles in amino acid and protein metabolism.

Low linoleic acid may facilitate Δ6 desaturase activity and docosahexaenoic acid accretion in human fetal development.

The n-3 and n-6 fatty acids are transferred across the placenta with consistently higher 22:6n-3 and lower 18:2n-6 in fetal than maternal plasma. This study sought to determine whether maternal and fetal cord blood red blood cell (RBC) phospholipid fatty acids show similar saturation with 22:6n-3, and also addressed the relationship between 18:2n-6 and Δ6 desaturase product/precursor ratios for 97 mothers and newborns. Despite higher fetal than maternal plasma phospholipid 22:6n-3, the maternal and fetal RBC phospholipid 22:6n-3 showed similar curvilinear relationships to the plasma phospholipid 22:6n-3. Risk of failure to achieve high RBC phospholipid 22:6n-3 increased sharply below a plasma phospholipid 22:6n-3 of 6.5g/100g fatty acids. Higher maternal and fetal 18:2n-6 was associated with lower RBC phospholipid 22:6n-3/22:5n-3, 22:5n-6/22:4n-6 and 18:3n-6/18:2n-6. These findings suggest low placental transfer of 18:2n-6 may be a specific mechanism to prevent inhibition of fetal Δ6 desaturase and facilitate fetal cellular phospholipid 22:6n-3 accretion.

Maternal vitamin D status in pregnancy and adverse pregnancy outcomes in a group at high risk for pre-eclampsia.

OBJECTIVE: To determine in a group of pregnant women if vitamin D status, based on serum 25-hydroxyvitamin D (25OHD) concentration, was associated with a subsequent risk of pre-eclampsia or adverse pregnancy outcomes. DESIGN: Prospective cohort study. SETTING: Vancouver, British Columbia, Canada (49°N). POPULATION: Women attending a specialist antenatal clinic because of clinical or biochemical risk factors for pre-eclampsia (n = 221). METHODS: Serum 25OHD concentration measured between 10 and 20 weeks of gestation. MAIN OUTCOME MEASURES: Pre-eclampsia and composite adverse pregnancy outcomes. RESULTS: Of the women, 78% were vitamin D insufficient (25OHD <75 nmol/l) and 53% were vitamin D deficient (25OHD <50 nmol/l). There was no difference in the rates of pre-eclampsia, gestational hypertension, preterm birth or composite adverse pregnancy outcomes by 25OHD concentration. CONCLUSIONS: Vitamin D deficiency and insufficiency were common in a group of women at high risk of pre-eclampsia; however, it was not associated with subsequent risk of an adverse pregnancy outcome.

Antioxidant capacity of human milk and its association with vitamins A and E and fatty acid composition.

AIM: The antioxidant capacity of human milk reflects the presence and activity of multiple components, which prevent oxidative rancidity. The aim of this study was to use the Oxygen Radical Absorbance Capacity assay to assess human milk antioxidant capacity and find correlations with milk components. METHODS: Milk samples collected from 60 breastfeeding women at 1 month postpartum were assayed for antioxidant capacity, vitamins E and A, and fatty acids. Potential statistical relationships of concentrations of vitamins A and E and polyunsaturated fatty acids on the antioxidant capacity of human milk were determined. RESULTS: Human milk antioxidant capacity was positively attributed to alpha-tocopherol concentration (rho < 0.05). The vitamin A concentration did not significantly contribute to milk antioxidant capacity, but was correlated to milk alpha-tocopherol concentration (r = 0.587; rho < 0.001). There was no evidence of an inverse relationship between polyunsaturated fatty acids concentration and the antioxidant capacity value of milk. CONCLUSION: This study shows that alpha-tocopherol is an important contributor to the oxidative stability of human milk. Moreover, there was no evidence obtained to show that women who have high levels of milk polyunsaturated fatty acids are predisposed to lower milk antioxidant capacity.

Are plasma homocysteine and methionine elevated when binging and purging behavior complicates anorexia nervosa? Evidence against the transdiagnostic theory of eating disorders.

OBJECTIVE: To determine whether plasma total homocysteine (tHcy) and plasma methionine levels are different in anorexia nervosa restricting type (AN-R) compared to anorexia nervosa binge eating/purging type (AN-BP). METHODS: Cross-sectional design. SUBJECTS: Subjects were recruited from the outpatient program of the Eating Disorders Program at St. Paul's Hospital, Vancouver, Canada. All subjects had a current Diagnostic and Statistical Manual of mental Disorders - Fourth Edition (DSM-IV) AN-R, or AN-BP diagnosis. Controls were recruited from staff and trainees of Child and Family Research Institute, and Children's and Women's Hospital, University of British Columbia. RESULTS: Samples were obtained from AN-R (N=30), AN-BP (N=32) and control women (N=73) and men (N=33). The 5- 95th% confidence intervals from the control women were taken as the normal range. The plasma tHcy and methionine for the control group had a 5-95th percentile range of 5.66-10.57 and 15.3-40.2 microM, respectively. Plasma tHcy was elevated in women with AN-BP (9.24+/-0.85 microM, N=32) but not with AN-R (7.90+/-0.38 microM, N=30). Plasma methionine was decreased in women with AN-BP (22.2+/-1.43 microM, N=32) compared to the control group of women (25.1+/-0.89 microM). The plasma methionine/tHcy ratio was elevated in the women with AN-BP (0.50+/-0.09) but not in those with AN-R (0.34+/-0.03). CONCLUSION: Elevated plasma tHcy and decreased plasma methionine are consistent with impaired homocysteine remethylation. Altered methyl transfer capacity or methyl deficiency could impair monoamine neurotransmitter metabolism potentially impacting cognitive and psychological function.We hypothesize that the treatment of AN-BP should consider the need for nutritional support of methyl metabolism.

Metformin influences cardiomyocyte cell death by pathways that are dependent and independent of caspase-3.

AIMS/HYPOTHESIS: Metformin has been shown to increase fatty acid oxidation, an effect mediated by AMP activated protein kinase (AMPK). We hypothesised that metformin could prevent both caspase-3 activation and apoptosis when induced by palmitic acid. MATERIALS AND METHODS: Cardiomyocytes were incubated with 1 mmol/l palmitic acid, in the absence or presence of metformin (1-5 mmol/l). Following 1 to 16 h, cell damage was evaluated by measuring lactate dehydrogenase released into the incubation medium, and Hoechst staining. To investigate the mechanism of metformin's effect on cardiomyocytes, substrate utilisation and phosphorylation of AMPK and acetyl-CoA carboxylase were measured. Intracellular mediators of apoptosis were also evaluated. RESULTS: Incubation of myocytes with palmitic acid for 16 h increased apoptosis, an effect that was partly blunted by 1 and 2 mmol/l metformin. This beneficial effect of metformin was associated with increased AMPK phosphorylation, palmitic acid oxidation and suppression of high-fat-induced increases in (1) long chain base biosynthesis protein 1 levels, (2) ceramide levels, and (3) caspase-3 activity. Unexpectedly, 5 mmol/l metformin dramatically increased apoptosis in myocytes incubated with high fat. This effect was associated with a robust increase in glycolysis, lactate accumulation, and a significant drop of pH in the myocyte incubation medium. CONCLUSIONS/INTERPRETATION: Our study demonstrates that metformin reduces high-fat-induced cardiac cell death, probably through inhibition of ceramide synthesis. However, at high concentrations, metformin causes proton and lactate accumulation, leading to cell damage that is independent of caspase-3.

Essential fatty acid transfer and fetal development.

Docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6) are important structural components of the central nervous system. These fatty acids are transferred across the placenta, and are accumulated in the brain and other organs during fetal development. Depletion of 22:6n-3 from the retina and brain results in reduced visual function and learning deficits: these may involve critical roles of 22:6n-3 in membrane-dependent signaling pathways and neurotransmitter metabolism. Transfer of 22:6n-3 across the placenta involves specific binding and transfer proteins that facilitate higher concentrations of 22:6n-3 and 20:4n-6, but lower linoleic acid (18:2n-6) in fetal compared with maternal plasma, or in the breast-fed or formula-fed infant. However, human and animal studies both demonstrate that maternal diet impacts fetal 22:6n-3 and 20:4n-6 accretion. After birth, parenteral lipid, human milk and infant formula feeding all result in a marked decrease in plasma 22:6n-3 and 20:4n-6 and an increase in 18:2n-6. Estimation of fetal tissue fatty acid accretion suggests that current preterm infant feeds are unlikely to meet in utero rates of 22:6n-3 accretion. Consideration needs to be given to whether fetal plasma 22:6n-3 and 20:4n-6 enrichment and the low 18:2n-6 facilitates accretion of 22:6n-3 and 20:4n-6 in developing tissues.

Are human milk long-chain polyunsaturated fatty acids related to visual and neural development in breast-fed term infants?

OBJECTIVE: To determine whether docosahexaenoic acid (DHA) is related to visual and neural development in term breast-fed infants. DESIGN: A prospective study of 83 infants who were exclusively breast-fed for at least 3 months. We determined red blood cell and plasma fatty acids at 2 months, visual acuity at 2, 4, 6, and 12 months, speech perception and an object search task at 9 months, Bayley's mental development index and psychomotor development index at 6 and 12 months, and novelty pReference at 6 and 9 months. RESULTS: The infant red blood cell phosphatidylethanolamine DHA was significantly related to visual acuity at 2 months of age (r = 0.32, P =.01) and 12 months of age (r = 0.30, P =.03). The ability to discriminate nonnative retroflex and phonetic contrasts at 9 months of age was related to the plasma phospholipid DHA (r = 0.48, P <.02) and red blood cell phosphatidylethanolamine DHA (r = 0.26, P =.02) at 2 months of age after adjusting for covariates. CONCLUSION: DHA may influence the development of visual acuity and neural pathways associated with the developmental progression of language acquisition in term breast-fed infants. The extent to which our results can be attributed solely to DHA from maternal sources through breast milk or in gestation or other confounding factors remains to be determined.

Growth and development in term infants fed long-chain polyunsaturated fatty acids: a double-masked, randomized, parallel, prospective, multivariate study.

OBJECTIVE: To evaluate the effects of dietary intake of the long-chain polyunsaturated fatty acids, arachidonic acid (AA), and docosahexaenoic acid (DHA) on multiple indices of infant growth and development. DESIGN: A double-masked, randomized, parallel trial was conducted with term infants fed formulas with or without AA+DHA for 1 year (N = 239). Reference groups of breastfed infants (N = 165) weaned to formulas with and without AA+DHA were also studied. Infants in the formula groups were randomized at

Anandamide and diet: inclusion of dietary arachidonate and docosahexaenoate leads to increased brain levels of the corresponding N-acylethanolamines in piglets.

Endogenous ligands of cannabinoid receptors have been discovered recently and include some N-acylethanolamines (NAEs; e.g., N-arachidonoylethanolamine) and some 2-acylglycerols (e.g., sn-2-arachidonoylglycerol). Previously, we found these compounds to be active biologically when administered per os in large quantities to mice. In the present work, piglets were fed diets with and without 20:4n-6 and 22:6n-3 fatty acid precursors of NAEs, in levels similar to those found in porcine milk, during the first 18 days of life, and corresponding brain NAEs were assessed. In piglets fed diets containing 20:4n-6 and 22:6n-3, there were increases in several biologically active NAEs in brain homogenates-20:4n-6 NAE (4-fold), 20:5n-3 NAE (5-fold), and 22:5n-3 and 22:6n-3 NAE (9- to 10-fold). These results support a mechanism we propose for dietary long-chain polyunsaturated fatty acids influences on brain biochemistry with presumed functional sequelae. This paradigm will enable targeted investigations to determine whether and why specific populations such as infants, elderly, or persons suffering from certain clinical conditions may benefit from dietary long-chain polyunsaturated fatty acids.

Infant plasma trans, n-6, and n-3 fatty acids and conjugated linoleic acids are related to maternal plasma fatty acids, length of gestation, and birth weight and length.

BACKGROUND: Arachidonic acid (AA) and docosahexaenoic acid (DHA) are important for growth and neural development. trans Fatty acids (TFAs) may inhibit desaturation of linoleic acid (LA) and alpha-linolenic acid (ALA) to AA and DHA, respectively. Conjugated linoleic acids (CLAs) also alter lipid metabolism and body fat. OBJECTIVE: We determined the associations of birth outcome with maternal and infant plasma concentrations of TFAs, CLAs, AA, and DHA. DESIGN: In healthy women, we sampled maternal blood at 35 wk gestation (n = 58) and umbilical cord blood at birth (n = 70). RESULTS: Mean (+/- SEM) TFA concentrations (% by wt) in infant plasma were as follows: triacylglycerol, 2.83 +/- 0.19 (range: 0.63-12.79); phospholipid, 0.67 +/- 0.03 (0.11-1.33); and cholesteryl ester, 2.04 +/- 0.01 (0.86-4.24). LA, AA, DHA, TFA, and CLA concentrations in infant phospholipids correlated with the same fatty acid in maternal plasma phospholipids (n = 44; P < 0.05). Infant plasma cholesteryl ester and triacylglycerol TFAs and cholesteryl ester CLAs (r = -0.33, -0.42, and -0.49, respectively) were significantly inversely related to length of gestation. Triacylglycerol and cholesteryl ester AA were positively related to length of gestation (r = 0.41 and 0.37, respectively) and birth weight (r = 0.27 and 0.23, respectively). Inverse correlations occurred between infant plasma TFA and DHA concentrations in triacylglycerols (r = -0.33) and between TFA and AA concentrations in cholesteryl esters (r = -0.23). CONCLUSION: The results suggest possible important effects of TFAs and of AA on fetal growth and length of gestation.

Cholesterol synthesis and accretion within various tissues of the fetal and neonatal rat.

The rate of cholesterol synthesis is reported to be higher in fetal relative to adult rats. Along with the observation that maternal diets high in fat and cholesterol are unable to alter the rate of cholesterol synthesis in the fetus, this has been taken as indirect evidence that the fetal rat meets its cholesterol needs through de novo synthesis. This study quantified the rates of cholesterol synthesis and accumulation in the liver, brain, intestine, and carcass of the fetal and neonatal rat and the placenta to determine whether these developing tissues are able to support their own cholesterol needs without the uptake of plasma lipoprotein cholesterol. The rate of cholesterol synthesis was determined in vivo using [3H]water. The rate of cholesterol accumulation was determined by calculating the difference in tissue cholesterol content between 2 subsequent days of development. Total fetal body cholesterol synthesis was sufficient to support the rate of cholesterol accumulation. Fetal and neonatal liver synthesized cholesterol at a rate in excess of cholesterol accumulation, suggesting hepatic secretion of cholesterol into the plasma. Before the onset of suckling, the rates of de novo cholesterol synthesis in the intestine, brain, and carcass were also sufficient but not higher than the need for cholesterol accretion. After the establishment of suckling, the rate of cholesterol accumulation in the intestine and carcass was in excess of synthesis, suggesting that neonatal tissues derive some of their cholesterol from dietary milk or liver. These studies suggest that the perinatal rat does not require exogenous cholesterol to support tissue cholesterol accretion. However, the fetal liver may support cholesterol accretion in other tissues through rates of synthesis in excess of accumulation and secretion into plasma. The placenta may derive some cholesterol from the maternal and/or fetal plasma.

Dietary fatty acid composition in pregnancy alters neurite membrane fatty acids and dopamine in newborn rat brain.

The importance of maternal dietary fatty acids on arachidonic acid [AA; 20:4(n-6)] and docosahexaenoic acid [DHA; 22:6(n-3)] in fetal brain nerve growth cone membranes and monoaminergic neurotransmitters was investigated. Rats were fed purified diets containing 20 g/100 g safflower oil with 74.3% 18:2(n-6), 0.2% 18:3(n-3), soybean oil with 55.4% 18:2(n-6), 7.7% 18:3(n-3) or high fish oil with 24.6% 22:6(n-3) through gestation. Tissue for rats within a litter were pooled at birth, brain growth cone membranes prepared and phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylinositol (PI) fatty acids quantified by gas-liquid chromatography. Dopamine, serotonin, and the metabolites 3,4-dihydroxyphenylacetic acid and homovanillic acid, and 5-hydroxyindolacetic acid were quantified by HPLC. Growth cone membranes from offspring of rats fed safflower oil had significantly lower, and offspring of rats fed high 22:6(n-3) fish oil had significantly higher 22:6(n-3) in PE, PS and PI than the soybean oil group. The growth cone membrane PC, PE and PS 20:4(n-6) was significantly lower in the fish oil than in the soybean or safflower oil groups. Serotonin concentration was significantly higher in brain of offspring in the safflower oil compared with the soybean oil group. The newborn brain dopamine was inversely related to PE DHA and PS DHA (P < 0.001), but positively related to PC AA (P < 0.05). These studies show that maternal dietary fatty acids may alter fetal brain growth cone (n-6) and (n-3) fatty acids, and neurotransmitters involved in neurite extension, target finding and synaptogenesis. The functional importance, however, is not known at this time.

The role of dietary n-6 and n-3 fatty acids in the developing brain.

The dietary requirements for essential fatty acids and the possibility of a specific role for the polyunsaturated fatty acid docosahexaenoic acid (DHA) is one of the most controversial areas in infant nutrition. DHA is found in unusually high concentrations in the brain and is selectively accumulated during fetal and infant brain growth. DHA can be synthesised through a complex series of chain elongation-desaturation reactions from alpha-linolenic acid, but the efficiency of this process in young infants is not clear. Clinical studies on the potential benefits to neural development of dietary DHA have yielded conflicting results. Recent studies have provided evidence that plasma DHA is available to developing brain and that DHA is involved in dopamine and serotonin metabolism. These findings should guide clinical studies to more sensitive measures of the functional roles of dietary n-3 fatty acids and to clinical conditions where n-3 fatty acids may have benefit.

Low erucic acid canola oil does not induce heart triglyceride accumulation in neonatal pigs fed formula.

Canola oil is not approved for use in infant formula largely because of concerns over possible accumulation of triglyceride in heart as a result of the small amounts of erucic acid (22:1n-9) in the oil. Therefore, the concentration and composition of heart triglyceride were determined in piglets fed from birth for 10 (n = 4-6) or 18 (n = 6) d with formula containing about 50% energy fat as 100% canola oil (0.5% 22:1n-9) or 100% soybean oil, or 26% canola oil or soy oil (blend) with palm, high-oleic sunflower and coconut oil, providing amounts of 16:0 and 18:1 closer to milk, or a mix of soy, high-oleic sunflower and flaxseed oils with C16 and C18 fatty acids similar to canola oil but without 22:1. Biochemical analysis found no differences in heart triglyceride concentrations among the groups at 10 or 18 d. Assessment of heart triglycerides using Oil Red O staining in select treatments confirmed no differences between 10-d-old piglets fed formula with 100% canola oil (n = 4), 100% soy oil (n = 4), or the soy oil blend (n = 2). Levels of 22:1n-9 in heart triglyceride and phospholipid, however, were higher (P<0.01) in piglets fed 100% canola oil or the canola oil blend, with higher levels found in triglycerides compared with phospholipids. The modest accumulation of 22:1n-9 associated with feeding canola oil was not associated with biochemical evidence of heart triglyceride accumulation at 10 and 18 d.

Diverse, region-specific effects of addition of arachidonic and docosahexanoic acids to formula with low or adequate linoleic and alpha-linolenic acids on piglet brain monoaminergic neurotransmitters.

Differences in visual, auditory, and learning tasks have been reported for infants and animals given diets varying in omega-3 fatty acids, but the neurobiochemical basis for these changes is unclear. This study investigated the effect of feeding formula with 0.8% energy C18:2omega-6 + 0.05% C18:3omega-3 (low), or 8.3% C18:2omega-6 + 0.8% C18:3omega-3 (adequate), with and without 0.2% energy arachidonic acid (C20:4omega-6) and 0.16% docosahexanoic acid (C22:6omega-3), on monoaminergic neurotransmitters in different brain regions of piglets fed formula from birth to 18 d. The amount of C18:2omega-6 + C18:3omega-3 fed in formula had a significant effect on frontal cortex dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, serotonin, and 5-hydroxyindolacetic acid; striatum serotonin and inferior colliculus serotonin, resulting in lower concentrations in piglets fed the low compared with adequate C18:2omega-6 + C18:3omega-3 formula. Inclusion of arachidonic acid and docosahexanoic acid in the low, but not in the adequate, C18:2omega-6 + C18:3omega-3 formula resulted in increased concentrations of all monoamines in the frontal cortex, and in striatum and inferior colliculus serotonin. Feeding arachidonic acid and docosahexanoic acid in the formulas increased dopamine and 5-hydroxyindolacetic acid in superior and inferior colliculus, areas related to processing and integration of visual and auditory information. Higher dopamine and 5-hydroxyindolacetic acid were found in these regions even when arachidonic acid and docosahexanoic acid were added to the C18:2omega-6 + C18:3omega-3 adequate formula. This study suggests that functional changes among animals and infants fed diets varying in omega-6 and omega-3 fatty acids may involve altered neurotransmitter metabolism.

Essential fatty acids in infant nutrition: lessons and limitations from animal studies in relation to studies on infant fatty acid requirements.

Animal studies have been of pivotal importance in advancing knowledge of the metabolism and roles of n-6 and n-3 fatty acids and the effects of specific dietary intakes on membrane composition and related functions. Advantages of animal studies include the rigid control of fatty acid and other nutrient intakes and the degree, timing, and duration of deficiency or excess, the absence of confounding environmental and clinical variables, and the tissue analysis and testing procedures that cannot be performed in human studies. However, differences among species in nutrient requirements and metabolism and the severity and duration of the dietary treatment must be considered before extrapolating results to humans. Studies in rodents and nonhuman primates fed diets severely deficient in alpha-linolenic acid (18:3n-3) showed altered visual function and behavioral problems, and played a fundamental role by identifying neural systems that may be sensitive to dietary n-3 fatty acid intakes; this information has assisted researchers in planning clinical studies. However, whereas animal studies have focused mainly on 18:3n-3 deficiency, there is considerable clinical interest in docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6) supplementation. Information from animal studies suggests that brain and retinal concentrations of 22:6n-3 plateau with 18:3n-3 intakes of approximately 0.7% of energy, but this requirement is influenced by dietary 18:2n-6 intake. Blood and tissue concentrations of 22:6n-3 increase as 22:6n-3 intake increases, with adverse effects on growth and function at high intakes. Animal studies can provide important information on the mechanisms of both beneficial and adverse effects and the pathways of brain 22:6n-3 uptake.

Dietary n-3 fatty acid restriction during gestation in rats: neuronal cell body and growth-cone fatty acids.

Growth cones are membrane-rich structures found at the distal end of growing axons and are the predecessors of the synaptic membranes of nerve endings. This study examined whether n-3 fatty acid restriction during gestation in rats alters the composition of growth cone and neuronal cell body membrane fatty acids in newborns. Female rats were fed a standard control diet containing soy oil (8% of fatty acids as 18:3n-3 by wt) or a semisynthetic n-3 fatty acid-deficient diet with safflower oil (0.3% of fatty acids as 18:3n-3 by wt) throughout normal pregnancy. Experiments were conducted on postnatal day 2 to minimize the potential for contamination from synaptic membranes and glial cells. Dietary n-3 fatty acid restriction resulted in lower docosahexaenoic acid (DHA) concentrations and a corresponding higher docosapentaenoic acid concentration in neuronal growth cones, but had no effects on neuronal cell body fatty acid concentrations. These studies suggest that accretion of DHA in growth cones, but not neuronal cell bodies, is affected by n-3 fatty acid restriction during gestation. Differences in other fatty acids or components between the semisynthetic and the standard diet, however, could have been involved in the effects on growth-cone DHA content. The results also provide evidence to suggest that the addition of new membrane fatty acids to neurons during development occurs along the shaft of the axon or at the growth cone, rather than originating at the cell body.