Placental Fatty Acid Transfer Research Articles

Alteration of erythrocyte membrane polyunsaturated fatty acids in preterm newborns with retinopathy of prematurity

Extremely preterm infants are at high risk for retinopathy of prematurity (ROP), a potentially blinding disease characterized by abnormalities in retinal vascularization. Whereas animal studies revealed that n-3 polyunsaturated fatty acids (PUFAs) may be of benefit in preventing ROP, human studies conducted on preterm infants during the 1st weeks of life showed no association between blood n-3 PUFA bioavailability and ROP incidence and/or severity, probably because of the influence of nutrition on the lipid status of infants. In the OmegaROP prospective cohort study, we characterized the erythrocyte concentrations of PUFAs in preterm infants aged less than 29 weeks gestational age (GA) without any nutritional influence. We show that GA is positively associated with the erythrocyte n-6 to n-3 PUFA ratio, and particularly with the ratio of arachidonic acid (AA) to docosahexaenoic acid (DHA), in infants with ROP. A time-dependent accumulation of AA at the expense of DHA seems to occur in utero in erythrocytes of preterm infants who will develop ROP, thus reinforcing previous data on the beneficial properties of DHA on this disease. In addition, preliminary data on maternal erythrocyte membrane lipid concentrations suggest modifications in placental transfer of fatty acids. Documenting the erythrocyte AA to DHA ratio at birth in larger cohorts might be useful to set up new prognostic factors for ROP.

Transferred maternal fatty acids stimulate fetal adipogenesis and lead to neonatal and adult obesity

The prevalence of adult and childhood obesity are increasing. Most of the human newborn’s body fat accumulates in the last half of intrauterine life. Fat in the fetus was thought to be mostly synthesized from glucose, but now it is commonly accepted that the bulk of it is the product of placental transfer of maternal fatty acids. Transported fatty acids originate in maternal plasma “free” fatty acids, fatty acids hydrolyzed from maternal plasma triglycerides, and the poly-unsaturated fatty acid component of maternal phospholipids. Glucose remains an important precursor of alpha-glycerol phosphate, to which most transported fatty acids are eventually esterified. Maternal plasma lipids are elevated in late pregnancy and even more in obese and diabetic pregnant women. This accelerates the placental transport of fatty acids. The hypothesis presented in this paper rests on the observations that the exponential increase in fat tissue in the human embryo’s body occurs in time to parallel the increase of lipids in the mother’s blood and depends on the chemical affinity of the transcription factor PPAR gamma to fatty acids and on fatty acid stimulation of adipocyte generation from precursor cells. The hypothesis asserts that transported maternal fatty acids activate the transcription factors in the fetus and initiate conversion of the mesenchymal stem cells into adipocytes. In obese and diabetic mothers, the higher plasma lipids facilitate increased placental fatty acid transfer. This will increase adipocyte generation and, through this, the prevalence of babies with increased fat cell size and number. Babies born with increased adipose tissue cellularity will have greater probability of growing up to become obese adolescents and adults. These newborns, whose obesity is hyperplastic as well as hypertrophic, as adults will have difficulty losing weight through diet and exercise or will regain the lost weight more quickly than others without these characteristics. Accordingly, increased placental fatty acid transfer and accelerated adipocyte generation may explain not only neonatal obesity, but some aspects of the adult obesity epidemic also. It is therefore recommended that prevention of fetal fat cell hyperplasia, by lowering maternal plasma lipids in mid and late pregnancy, should be attempted in pregnancies at risk for macrosomia.

Placental fatty acid transfer.

This review outlines recent advances in placental lipid transport in relation to maternal metabolic status and pregnancy outcome. A particular focus of this review will be on the way these findings may influence our understanding of placental transfer of the essential fatty acid docosahexaenoic acid (DHA) which is crucial for fetal neurodevelopment and of lipid transfer as a predisposing factor for childhood obesity. Placental metabolism may determine the quantity and composition of fatty acids delivered to the fetus. Maternal factors, such as obesity, appear to regulate placental lipid metabolism and may influence fatty acids delivery to the fetus. Although the role of placental metabolism is now recognized, new evidence also suggests important roles for nontraditional fatty acid transporters such as Mfsd2a which facilitates transfer of DHA. Placental lipid metabolism is likely to be a determinant of placental transfer of fatty acids to the fetus. Maternal conditions, such as obesity, have now been shown to regulate placental lipid metabolism and thus may influence fatty acid transfer and fetal development. However, it is not yet clear how regulation of placental lipid metabolism affects fatty acid delivery to the fetus and its long-term health.

Milk-fat globule epidermal growth factor 8 (MFGE8) is expressed at the embryo- and fetal-maternal interface in equine pregnancy.

Milk-fat globule epidermal growth factor (EGF) 8 protein (MFGE8), also known as lactadherin, promotes cell adhesion in an Arg-Gly-Asp (RGD)-dependent modus via integrins. In the present study, the expression of MFGE8 was examined in equine endometrium during oestrus and at Days 12 and 16 after ovulation in pregnant and non-pregnant mares and in mares during the 5th month of gestation. Results demonstrated that MFGE8 is expressed at the embryo- and fetal-maternal interface in equine pregnancy. In non-pregnant endometrium its expression was upregulated by oestrogen, a finding that was confirmed using endometrial explant culture. MFGE8 was expressed at similar levels by conceptuses collected 13 and 14 days after ovulation and by allantochorion sampled during the 5th month of gestation. Pericytes of endometrial blood vessels displayed strong MFGE8 expression upon in situ hybridisation. During the 5th month of gestation, the fetal side of the allantochorionic villi in particular displayed pronounced staining upon in situ hybridisation, confirming that MFGE8 expression is not restricted to early pregnancy but persists and is present at the fetal-maternal interface. Potential roles of MFGE8 in equine pregnancy include mediating cell-cell adhesion, promotion of angiogenesis and placental transfer of fatty acids.

Placental dimethyl acetal fatty acid derivatives are elevated in preeclampsia

Preeclampsia (PE) was shown to affect the placental content and the transfer of polyunsaturated fatty acids (PUFA) to the fetus. Plasmalogens, a type of phospholipids with a vinyl-ether link at the sn-1 position, play an antioxidant role and are specifically enriched in PUFA at the sn-2 position. In this study, we characterized plasmalogen-derived dimethyl acetal (DMA) fatty acid derivatives, 16:0 DMA, 18:0 DMA, 9c-/11c-18:1 DMA and PUFA in the placenta of normotensive (n = 20) and PE (n = 20) pregnancies, according to the sampling site: peri-insertion or periphery. Phospholipid fatty acids from the placenta and maternal erythrocytes were identified by gas chromatography mass spectrometry and quantified by flame ionization detection. We found elevated total DMA in the PE placenta by 18% when compared to normotensive controls (p = 0.026). Moreover, the 16:0 DMA account for more than 55% of DMA fatty acids measured in the placenta, and its level is significantly higher in PE than controls (p = 0.018). Also, we found elevated placental PUFA, 20:5(n-3), 22:5(n-3) and a low level of 20:4(n-3) in PE compared to controls. Placental DMA was highly correlated with n-6 and n-3 PUFA in both, normotensive and PE pregnancies. In sum, elevated DMA fatty acids in the PE placenta could be an indirect defensive mechanism against oxidative stress and poor placental fatty acid transfer in PE.

Fate of orally administered radioactive fatty acids in the late-pregnant rat.

To investigate the biodisponibility of placental transfer of fatty acids, rats pregnant for 20 days were given tracer amounts of [(14)C]palmitic (PA), oleic (OA), linoleic (LA), α-linolenic (LNA), or docosahexaenoic acid (DHA) orally and euthanized at 0.5, 1.0, 2.0, or 8.0 h thereafter. Maternal plasma radioactivity in lipids initially increased only to decline at later times. Most of the label appeared first as triacylglycerols (TAG); later, the proportion in phospholipids (PhL) increased. The percentage of label in placental lipids was also always highest shortly after administration and declined later; again, PhL increased with time. Fetal plasma radioactivity increased with time, with its highest value at 8.0 h after DHA or LNA administration. DHA initially appeared primarily in the nonesterified fatty acids (NEFA) and PA, OA, LA, and LNA as TAG followed by NEFA; in all cases, there was an increase in PhL at later times. Measurement of fatty acid concentrations allowed calculation of specific (radio)activities, and the ratio (fetal/maternal) of these in the plasmas gave an index of placental transfer activity, which was LNA > LA > DHA = OA > PA. It is proposed that a considerable proportion of most fatty acids transferred through the placenta are released into the fetal circulation in the form of TAG.

Computational modelling of fatty acid transport in the human placenta.

Fatty acids are critical for normal fetal growth and development. The placenta mediates the transfer of fatty acids from the maternal to the fetal circulation. Yet, the mechanisms of fatty acid transport are not fully understood. The development of a computational model alongside experiments will test our understanding of the transfer mechanisms. Modelling experimental data suggest the presence of a metabolic pool within placental tissue that could represent the rate-limiting factor for fatty acid transfer. In addition the model suggests a slower flux capacity of the fetal-side of the placenta compared with the maternal-side. The model provides key insights into placental fatty acid transfer which will form the basis for future experimentation.

Placental Fatty Acid Transfer: A Key Factor in Fetal Growth

The functionality of the placenta may affect neonatal adiposity and fetal levels of key nutrients such as long-chain polyunsaturated fatty acids. Fetal macrosomia and its complications may occur even in adequately controlled gestational diabetic (GDM) mothers, suggesting that maternal glycemia is not the only determinant of fetal glycemic status and wellbeing. We studied in vivo the placental transfer of fatty acids (FA) labeled with stable isotopes administered to 11 control and 9 GDM pregnant women (6 treated with insulin). Subjects received orally ¹³C-palmitic, ¹³C-oleic, and ¹³C-linoleic acids and ¹³C-docosahexaenoic acid (¹³C-DHA) 12 h before an elective caesarean section. FA were quantified by gas chromatography and ¹³C enrichments by gas chromatography-isotope ratio mass spectrometry. The ¹³C-FA concentration was higher in total lipids of maternal plasma in GDM patients versus controls, except for ¹³C-DHA. Moreover, ¹³C-DHA showed a lower placenta/maternal plasma ratio in GDM patients versus controls and a significantly lower cord/maternal plasma ratio. Other FA ratios studied were not different between GDM and controls. A disturbed ¹³C-DHA placental uptake occurred in GDM patients treated with diet or insulin, while the latter also had lower ¹³C-DHA levels in the venous cord. The tracer study pointed towards an impaired placental DHA uptake as a critical step, while the transfer of other ¹³C-FA was less affected. Patients with GDM treated with insulin could also have a greater fetal fat storage, which may have contributed to the reduced ¹³C-DHA in the venous cord observed. The DHA transfer to the fetus was reduced in GDM pregnancies compared to controls. This might have an influence on fetal neurodevelopment and long-term consequences for the child.

Materno-fetal transfer of docosahexaenoic acid is impaired by gestational diabetes mellitus

Better knowledge on the disturbed mechanisms implicated in materno-fetal long-chain polyunsaturated fatty acid (LC-PUFA) transfer in pregnancies with gestational diabetes mellitus (GDM) may have potentially high implications for later on in effective LC-PUFA supplementation. We studied in vivo placental transfer of fatty acids (FA) using stable isotope tracers administrated to 11 control and 9 GDM pregnant women (6 treated with insulin). Subjects received orally [(13)C]palmitic, [(13)C]oleic and [(13)C]linoleic acids, and [(13)C]docosahexaenoic acid ((13)C-DHA) 12 h before elective caesarean section. Maternal blood samples were collected at -12, -3, -2, and -1 h, delivery, and +1 h. Placental tissue and venous cord blood were also collected. FA were quantified by gas chromatography (GC) and (13)C enrichments by GC-isotope ratio mass spectrometry. [(13)C]FA concentration was higher in total lipids of maternal plasma in GDM vs. controls, except for [(13)C]DHA. Moreover, [(13)C]DHA showed lower placenta/maternal plasma ratio in GDM vs. controls and significantly lower cord/maternal plasma ratio. For the other studied FA, ratios were not different between GDM and controls. Disturbed [(13)C]DHA placental uptake occurs in both GDM treated with diet or insulin, whereas the last ones also have lower [(13)C]DHA in venous cord. The tracer study pointed toward impaired placental DHA uptake as critical step, whereas the transfer of the rest of [(13)C]FA was less affected. GDM under insulin treatment could also have higher fetal fat storage, contributing to reduce [(13)C]DHA in venous cord. DHA transfer to the fetus was reduced in GDM pregnancies compared with controls, which might affect the programming of neurodevelopment in their neonates.

Race, pregravid BMI and gestational weight gain determine placental fatty acid transfer

Long chain polyunsaturated fatty acids (LCPUFA), arachidonic acid (AA) and docosoahexaenoic acid (DHA), are essential fatty acids for fetal growth (AA) and development (DHA). The fetus relies on maternal stores and placental transfer for LCPUFAs. In our study population of Hispanic and African‐American women, the relationships between maternal and fetal red blood cell (RBC) AA and DHA were examined and predictors of fetal fatty acids such as age, maternal pregavid body mass index (BMI), gestational weight gain, and race were determined. Maternal and venous cord blood and health histories were obtained at delivery (n=9). Gas chromatography was used to assess RBC fatty acids (wt %). Pearson correlations were used to analyze relationships. Stepwise regression determined the most significant predictors of AA and DHA transfer. There was preferential transfer of DHA and AA from mother to fetus over precursor fatty acids. Maternal DHA was positively correlated with fetal DHA (p=0.046), while AA was not. BMI and gestational weight gain were the strongest predictors of AA transfer to the fetus (p=0.045). DHA was less efficiently transferred in Hispanics than in African‐Americans (p=0.036). Future work is needed to better understand how other factors such as maternal diet affect these findings.Funded by a Howard Hughes Medical Institute grant to LSU (ARS); LSU AgCenter; USDA‐AFRI (CJL‐K)

Placental transfer of fatty acids and fetal implications

Considerable amounts of long-chain polyunsaturated fatty acids (LC-PUFAs), particularly arachidonic acid and docosahexaenoic acid (DHA, 22:6n-3), are deposited in fetal tissues during pregnancy; and this process is facilitated by placental delivery. Nevertheless, the mechanisms involved in LC-PUFA placental transfer remain unclear. Stable isotope techniques have been used to study human placental fatty acid transfer in vivo. These studies have shown a significantly higher ratio of (13)C-DHA in cord to maternal plasma compared with other fatty acids, which reflects a higher placental DHA transfer. In addition, a selective DHA accumulation in placental tissue, relative to other fatty acids, has been reported. The materno-fetal transfer of fatty acids is a slow process that requires ≥12 h. A high incorporation of dietary (13)C-DHA into maternal plasma phospholipids appears to be important for placental uptake and transfer. DHA in cord blood lipids correlates with placental messenger RNA expression of fatty acid transport protein (FATP)-4, compatible with a role of FATP-4 in DHA transfer. Impaired materno-fetal LC-PUFA transport has been proposed in pregnancies complicated by abnormal placental function (eg, due to gestational diabetes mellitus or intrauterine growth restriction), which should be addressed in future studies. Given that placental DHA transfer is important for child outcomes, elucidation of its potential modulation by transport mechanisms, maternal diet, and disease appears to be important.

Mechanisms involved in the selective transfer of long chain polyunsaturated fatty acids to the fetus

The concentration of long chain polyunsaturated fatty acid (LCPUFA) in the fetal brain increases dramatically from the third trimester until 18 months of life. Several studies have shown an association between the percentage of maternal plasma docosahexaenoic acid (DHA) during gestation and development of cognitive functions in the neonate. Since only very low levels of LCPUFA are synthesized in the fetus and placenta, their primary source for the fetus is the maternal circulation. Both in vitro and human in vivo studies using labeled fatty acids have shown preferential transfer of LCPUFA from the placenta to the fetus compared with other fatty acids, although the mechanisms involved are still uncertain. The placenta takes up circulating maternal non-esterified fatty acids (NEFA) and fatty acids released mainly by maternal lipoprotein lipase and endothelial lipase. These NEFA may enter the cell by passive diffusion or by means of membrane carrier proteins. Once in the cytosol, NEFA bind to cytosolic fatty acid-binding proteins for transfer to the fetal circulation or can be oxidized within the trophoblasts, and even re-esterified and stored in lipid droplets. Although trophoblast cells are not specialized for lipid storage, LCPUFA may up-regulate peroxisome proliferator activated receptor-γ (PPARγ) and hence the gene expression of fatty acid transport carriers, fatty acid acyl-CoA-synthetases and adipophilin or other enzymes involved in lipolysis, modifying the rate of placental transfer, and metabolism. The placental transfer of LCPUFA during pregnancy seems to be a key factor in the neurological development of the fetus. Increased knowledge of the factors that modify placental transfer of fatty acids would contribute to our understanding of this complex process.

DOCOSAHEXAENOIC ACID SUPPLEMENTATION DURING PREGNANCY MODULATES THE GENE EXPRESSION OF SOME FATTY ACID TRANSPORT PROTEINS IN HUMAN PLACENTA

Introduction: The placental transfer of fatty acids is a complex process mediated by specific carrier proteins from membranes and cytosol. Nevertheless, the underlying mechanisms for the preferential materno-fetal transfer of long-chain polyunsaturated fatty acids (LC-PUFA) are still poorly understood. The aim of this study is to evaluate the effects of nutritional supplements with docosahexaenoic acid (DHA) and folic acid on the placental mRNA expression of proteins that may regulate fatty acid transfer. Methods: We recruited 137 pregnant women from Granada (Spain) who received from the week 20 until delivery different nutritional supplements: placebo; MTHF supplemented with 400 ug folic acid; DHA supplemented with 500 mg DHA + 150 mg EPA; and DHA + MTHF (400ug folic acid, 500 mg DHA, 150 mg EPA ). Placental tissue was sampled at birth, fatty acid profile analyzed by gas-liquid chromatography, and mRNA expression of placental fatty acid transport proteins-1 (FATP-1) and-4 (FATP-4), heart- fatty acid binding protein (H-FABP) by real time-PCR. Results: No significant difference was found in the gene expression of fatty acid transport proteins FATP-1, FATP-4 or H-FABP among the 4 groups. There was a positive correlation of DHA % in placental phospholipids with mRNA expression of membrane proteins FATP-1 (R = +0.364, P = 0.001) and FATP-4 (R = +0.387, P < 0.001). Arachidonic acid/DHA ratio (AA/DHA) was inversely correlated with FATP-1 and FATP-4, respectively, in all placental lipid fractions except for cholesterol esters (phospholipids: R = −0.287, P = 0.007; R = −0.335, P = 0.002; NEFA: R = −0.213, P = 0.005; R = −0.235, P = 0.032; triglycerides: R= −0.310, P = 0.004; R = −0.407, P < 0.001). In contrast, the cytosolic protein H-FABP was not correlated with LC-PUFA. Conclusion: In uncomplicated pregnancies the mRNA expressions of membrane placental proteins FATP-1 and FATP-4 are related to placental accumulation of DHA and AA, respectively, whereas there is no correlation with expression of the cytosolic protein H-FABP.

THE INFLUENCE OF HABITUAL DIET AND INCREASED DOCOSAHEXAENOIC ACID INTAKE DURING PREGNANCY ON THE FATTY ACID COMPOSITION OF INDIVIDUAL PLACENTAL LIPIDS

Aim: To study the effects of dietary n-3 fatty acids on the composition of individual placenta lipid fractions and elucidate the involvement of various lipid fraction in placental fatty acid transfer. Methods: We analysed the fatty acid composition of placental phospholipids (PL), triglycerides (TG), cholesterol esters (CE) and non-esterified fatty acids (NEFA) in pregnant women receiving placebo or n-3 long-chain polyunsaturated fatty acids. Placental samples were available from 202 women participating in a randomised clinical trial in Germany, Spain and Hungary. During the second half of pregnancy, the women received milk based supplements containing either modified fish oil [500 mg docosahexaenoic acid (DHA), 150 mg eicosapentaenoic acid daily] or placebo. Placental samples were collected immediately after delivery, lipids extracted, lipid fractions isolated by thin layer chromatography and fatty acids analysed by gas chromatography. Results: DHA values (M ± SE) of placebo groups tended to differ between countries (table). Relative to placebo, supplementation increased DHA content (M ± SE) of all placental lipid fractions in all study populations.TableTotal saturated and monounsaturated fatty acids were not affected by the supplementation, but n-6 long chain polyunsaturated fatty acids tended to be lower after supplementation (n.s.). There was a significant correlation of DHA content in TG (r = 0.55) and PL (r = 0.71) to DHA content in NEFA. Conclusions: Dietary n-3 fatty acids modifies the composition of all major lipid fractions in placental tissue. The correlation of DHA contents in NEFA with those of TG and PL, respectively, are in line with a placental NEFA uptake and subsequent incorporation into tissue lipid fractions.

PC63 EXPRESSION PATTERN OF FATTY ACID TRANSPORT PROTEIN-1 (FATP-1), -4 (FATP-4), AND HEART-FATTY ACID BINDING PROTEIN (H-FABP) GENES IN HUMAN TERM PLACENTA

Introduction: Placental fatty acid transfer is critical to meet the fetal requirements for biosynthesis of biological membranes, myelin and various signaling molecules. Increasing evidence suggests that the direction and magnitude of fatty acid flux is mainly determined by the availability of transfer proteins (Dutta-Roy, 2000). However, the relative expression of these proteins in human placental tissue is not known. The aim of this study was to elucidate the human placental mRNA expression of proteins that may regulate placental fatty acid transfer. Methods: We collected placental tissue from five pregnant women undergoing caesarean section at 38+/−1.1 weeks of gestation and from five different placenta locations. We quantified the fatty acid composition at the individual locations by gas-liquid chromatography (Klingler et al., 2003) and mRNA expression of placental fatty acid transport proteins-1 (FATP-1) and −4 (FATP-4), heart-fatty acid binding protein (H-FABP) and liver-FABP (L-FABP) by real time-PCR. The mRNA expression of these proteins was expressed relative to that of the house keeping gene beta-2-microglobulin. Results: The mRNA expression of plasma membrane proteins FATP-1 and FATP-4 (39.5+/−6.4a, and 16.4+/−2.8b) was significantly higher than that of the cytosolic proteins H-FABP (9.3+/−0.7c) and L-FABP. The expression of L-FABP was too low in placenta for accurate quantification. No significant differences were detected in the expression of FATP-1, FATP-4 and H-FABP between the different placental locations. Conclusion: Our results indicate the feasibility of reproducible quantification of placental mRNA expression of some fatty acid transport proteins and provide first results on their expression in human placentas at term. Although a higher expression of membrane bound transport proteins than of cytosolic transport proteins was demonstrated, further studies are needed to establish the role of the individual placental proteins in the fetal accretion of fatty acids and their response to maternal diet and disease.

In vivo investigation of the placental transfer of 13C-labeled fatty acids in humans

Placental fatty acid transfer in humans in vivo was studied using stable isotopes. Four pregnant women undergoing cesarean section received 4 h before delivery an oral dose of [(13)C]palmitic acid (PA), [(13)C]oleic acid (OA), [(13)C]linoleic acid (LA), and [(13)C]docosahexaenoic acid (DHA). Maternal blood samples were collected at -4 h (basal), -3 h, -2 h, -1 h, 0 h, and +1 h relative to time of cesarean section. At the time of birth, venous cord blood and placental tissue were collected. Fatty acid composition was determined by gas-liquid chromatography and isotopic enrichment by gas chromatography-combustion-isotope ratio mass spectrometry. (13)C-enrichment of fatty acids in the nonesterified fatty acids (NEFA) of cord plasma tended to be higher than in NEFA of placenta, with statistically significant differences for the nonesterified OA and DHA ([(13)C]PA, 0.024 +/- 0.011 vs. 0.001 +/- 0.001; [(13)C]OA, 0.042 +/- 0.008 vs. 0.005 +/- 0.003; [(13)C]LA, 0.038 +/- 0.010 vs. 0.008 +/- 0.002; [(13)C]DHA, 0.059 +/- 0.009 vs. 0.010 +/- 0.003). The ratio of tracer fatty acid concentrations of placenta to maternal plasma was significantly higher for [(13)C]DHA than for the other fatty acids ([(13)C]PA, 7.1 +/- 1%; [(13)C]OA, 3.8 +/- 0.4%; [(13)C]LA, 9.2 +/- 1.3%; [(13)C]DHA, 25.9 +/- 3.4%). These results suggest that only a part of the placental NEFA participated in fatty acid transfer, and that the placenta showed a preferential accretion of DHA relative to the other fatty acids.

Expression of PPAR and RXR Isoforms in the Developing Rat and Human Term Placentas

Placental fatty acid transfer is critical to meet the foetal requirements necessary for the biosynthesis of biological membranes, myelin, and various signaling molecules. The primary objective of this research was to elucidate the placental expression patterns of genes that may potentially regulate placental fatty acid transfer and homeostasis. In this study, we have elucidated the temporal and spatial patterns of expression of peroxisome proliferator-activated receptor (PPAR) and 9-cis retinoic acid receptor (RXR) isoforms in the junctional and labyrinth zones of the developing rat chorioallantoic placenta and in human term placenta. PPAR (α, β, and γ) and RXR (α, β, and γ) isoforms are nuclear hormone receptors that are known to regulate gene transcription and protein expression levels of fatty acid transport and metabolism mediating proteins through the formation of a DNA binding heterodimer complex. In the present study, the expression patterns of PPAR and RXR isoforms were determined in developing rat placenta and human term placenta using RT-PCR and immunohistochemical analyses. PPARα, β, γ, RXRα, β and γ were expressed in both junctional (invasive/endocrine function) and labyrinth (transport barrier) zones of the rat placenta, from day 13 to day 21 of gestation. In the human term placenta, PPARα, β, γ, RXRα and γ were observed, while RXRβ was not detected. Immunocytochemistry staining results determined the presence of PPARα, β, γ, RXRα and γ to be specific to the syncytial trophoblast layer of the human chorionic villi. The presence of PPAR and RXR isoforms in both the rat and human placentas suggest that PPAR and RXR isoforms are potential regulators of placental lipid transfer and homeostasis. Our work provides a framework for the further investigation of PPAR and RXR isoform specific regulation of placental fatty acid uptake, transport and metabolism.

Lipid metabolism in the fetus and the newborn.

During late gestation, although maternal adipose tissue lipolytic activity becomes enhanced, lipolytic products cross the placenta with difficulty. Under fasting conditions, free fatty acids (FFA) are used for ketogenesis by the mother, and ketone bodies are used as fuels and lipogenic substrates by the fetus. Maternal glycerol is preferentially used for glucose synthesis, saving other gluconeogenic substrates, like amino acids, for fetal growth. Placental transfer of triglycerides is null, but essential fatty acids derived from maternal diet, which are transported as triglycerides in lipoproteins, become available to the fetus owing to the presence of both lipoprotein receptors and lipase activities in the placenta. Diabetes in pregnancy promotes lipid transfer to the fetus by increasing the maternal-fetal gradient, which may contribute to an increase in body fat mass in newborns of diabetic women. Deposition of fat stores in the fetus is very low in the rat but high in humans, where body fat accretion occurs essentially during the last trimester of intra-uterine life. This is sustained by the intense placental transfer of glucose and by its use as a lipogenic substrate, as well as by the placental transfer of fatty acids and to their low oxidation activity. During the perinatal period an active ketonemia develops, which is maintained in the suckling newborn by several factors: (i) the high-fat and low-carbohydrate content in milk, (ii) the enhanced lipolytic activity occurring during the first few hours of life, and (iii) both the uptake of circulating triglycerides by the liver due to the induction of lipoprotein lipase (LPL) activity in this organ, and the presence of ketogenic activity in the intestinal mucose. Changes in LPL activity, lipogenesis and lipolysis contribute to the sequential steps of adipocyte hyperplasia and hypertrophia occurring during the extra-uterine white adipose tissue development in rat, and this may be used as a model to extrapolate the intra-uterine adipose tissue development in other species, including humans.

Lipid metabolism in the fetus and the newborn

During late gestation, although maternal adipose tissue lipolytic activity becomes enhanced, lipolytic products cross the placenta with difficulty. Under fasting conditions, free fatty acids (FFA) are used for ketogenesis by the mother, and ketone bodies are used as fuels and lipogenic substrates by the fetus. Maternal glycerol is preferentially used for glucose synthesis, saving other gluconeogenic substrates, like amino acids, for fetal growth. Placental transfer of triglycerides is null, but essential fatty acids derived from maternal diet, which are transported as triglycerides in lipoproteins, become available to the fetus owing to the presence of both lipoprotein receptors and lipase activities in the placenta. Diabetes in pregnancy promotes lipid transfer to the fetus by increasing the maternal-fetal gradient, which may contribute to an increase in body fat mass in newborns of diabetic women. Deposition of fat stores in the fetus is very low in the rat but high in humans, where body fat accretion occurs essentially during the last trimester of intra-uterine life. This is sustained by the intense placental transfer of glucose and by its use as a lipogenic substrate, as well as by the placental transfer of fatty acids and to their low oxidation activity. During the perinatal period an active ketonemia develops, which is maintained in the suckling newborn by several factors: (i) the high-fat and low-carbohydrate content in milk, (ii) the enhanced lipolytic activity occurring during the first few hours of life, and (iii) both the uptake of circulating triglycerides by the liver due to the induction of lipoprotein lipase (LPL) activity in this organ, and the presence of ketogenic activity in the intestinal mucose. Changes in LPL activity, lipogenesis and lipolysis contribute to the sequential steps of adipocyte hyperplasia and hypertrophia occurring during the extra-uterine white adipose tissue development in rat, and this may be used as a model to extrapolate the intra-uterine adipose tissue development in other species, including humans.

Effects of various dietary fats on cardiolipin acyl composition during ontogeny of mice.

Cardiolipin (CL) is a unique mitochondrial phospholipid, containing up to 85 wt% 18:2n-6 in mammals. The influence of maternal dietary fatty acids on the acyl composition of offspring CL has not been examined previously. Adult female mice were thus fed diets rich in 18:1n-9 (olive oil), 18:2n-6 (safflower oil), 18:3n-3 (linseed oil) or 20:5n-3 and 22:6n-3 (fish oil/safflower, 9:1, w/w), for a five month period, encompassing two breeding cycles. Offspring from the second breeding cycle were then fed these diets. The acyl composition of CL, phosphatidylcholine and phosphatidylethanolamine from liver and heart was evaluated from mice killed 3, 18 and 42 days after parturition. The primary nutrient sources at these three time points were transplacental nutrients, breast milk and the diet, respectively. Maternal diet was found to influence the acyl composition of CL via both placental transfer of fatty acids and breast milk. Fish oil feeding resulted in replacement of a substantial portion of 18:2n-6 with 22:6n-3; after 42 days, the area% of 18:2n-6 in heart CL was reduced from 62% in safflower oil fed mice to 12%. In comparison to fish oil feeding, linseed oil feeding resulted in a much lower accumulation of 22:6n-3. Olive oil feeding resulted in substantial replacement of 18:2n-6 with 18:1n-9 (18:2n-6 was reduced from 62% to 31%). Physiologically, these findings are relevant because changes in CL acyl composition may influence the activity of associated inner mitochondrial membrane enzymes.