Transfer Of Polyunsaturated Fatty Acids Research Articles

Zooplankton in northern lakes show taxon‐specific responses in fatty acids across climate‐productivity and ecosystem size gradients

AbstractNorthern lakes are facing rapid environmental alterations—including warming, browning, and/or changes in nutrient concentrations—driven by climate change. These environmental changes can have profound impacts on the synthesis and trophic transfer of polyunsaturated fatty acids (PUFA), which are important biochemical molecules for consumer growth and reproduction. Zooplankton are a key trophic link between phytoplankton and fish, but their biochemical responses to environmental change are not well understood. In this study, we assess the trends in fatty acid (FA) composition of zooplankton taxa among 32 subarctic and temperate lakes across broad climate‐productivity and ecosystem size gradients. We found that genus‐level taxonomy explained most FA variability in zooplankton (54%), suggesting that environmental changes that alter the taxonomic composition also affect the FA composition of zooplankton communities. Furthermore, the FA responses and their underlying environmental drivers differed between cladocerans and copepods. Cladocerans, including widespread Bosmina spp. and Daphnia spp., showed pronounced responses across the climate‐productivity gradient, with abrupt declines in PUFA, particularly eicosapentaenoic acid and arachidonic acid in warmer, browner, and more eutrophic lakes. Conversely, calanoid copepods had high and relatively stable PUFA levels across the gradient. In addition, all zooplankton taxa increased in stearidonic acid levels in larger lakes where PUFA‐rich cryptophytes were more abundant. Overall, our results suggest that climate‐driven environmental alterations pose heterogeneous impacts on PUFA levels among zooplankton taxa, and that the negative impacts of climate warming are stronger for cladocerans, especially so in small lakes.

The Role of the Grass Snake Natrix natrix (Reptilia, Colubridae) in the Transfer of Polyunsaturated Fatty Acids from Aquatic to Terrestrial Ecosystems to Land

As a result of analyses of fatty acid (FA) composition in the grass snake Natrix natrix and its food objects, tadpoles and metamorphs of two amphibian species: the moor frog Rana arvalis and the Pallas spadefoot toad Pelobates vespertinus, it was shown for the first time that the high total content of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids in the biomass of the snakes indicates its important role in the transfer of these essential substances from aquatic ecosystems to land. It was found that, since food sources of DHA in terrestrial ecosystems are absent, its high level in R. arvalis metamorps and grass snakes may be provided only by synthesis from biochemical precursors contained in food of aquatic origin.

Trophic transfer of polyunsaturated fatty acids across the aquatic-terrestrial interface: An experimental tritrophic food chain approach.

Aquatic and their adjacent terrestrial ecosystems are linked via the flux of organic and inorganic matter. Emergent aquatic insects are recognized as high-quality food for terrestrial predators, because they provide more physiologically relevant long-chain polyunsaturated fatty acids (PUFA) than terrestrial insects. The effects of dietary PUFA on terrestrial predators have been explored mainly in feeding trials conducted under controlled laboratory conditions, hampering the assessment of the ecological relevance of dietary PUFA deficiencies under field conditions. We assessed the PUFA transfer across the aquatic-terrestrial interface and the consequences for terrestrial riparian predators in two outdoor microcosm experiments. We established simplified tritrophic food chains, consisting of one of four basic food sources, an intermediary collector gatherer (Chironomus riparius, Chironomidae), and a riparian web-building spider (Tetragnatha sp.). The four basic food sources (algae, conditioned leaves, oatmeal, and fish food) differed in PUFA profiles and were used to track the trophic transfer of single PUFA along the food chain and to assess their potential effects on spiders, that is, on fresh weight, body condition (size-controlled measurement of nutritional status), and immune response. The PUFA profiles of the basic food sources, C.riparius and spiders differed between treatments, except for spiders in the second experiment. The PUFA α-linolenic acid (ALA, 18:3n-3) and ɣ-linolenic acid (GLA, 18:3n-6) were major contributors to the differences between treatments. PUFA profiles of the basic food sources influenced the fresh weight and body condition of spiders in the first experiment, but not in the second experiment, and did not affect the immune response, growth rate, and dry weight in both experiments. Furthermore, our results indicate that the examined responses are dependent on temperature. Future studies including anthropogenic stressors would deepen our understanding of the transfer and role of PUFA in ecosystems.

Lipid turnover and SQUAMOSA promoter-binding proteins mediate variation in fatty acid desaturation under early nitrogen deprivation revealed by lipidomic and transcriptomic analyses in Chlorella pyrenoidosa.

Nitrogen deprivation induces variations in fatty acid desaturation in microalgae, which determines the performance of biodiesel and the nutritional value of bioproducts. However, the detailed scenario and the underlying regulatory mechanism remain unclear. In this study, we attempt to outline these scenario and mechanisms by performing biochemical, lipidomic, and transcriptomic analyses in Chlorella pyrenoidosa and functional characterization of transcription factors in Yarrowia lipolytica. We found that early nitrogen deprivation dramatically reduced fatty acid desaturation without increasing lipid content. The contents of palmitic acid (16:0) and oleic acid (18:1) dramatically increased to 2.14 and 2.87 times that of nitrogen repletion on the second day, respectively. Lipidomic analysis showed the transfer of polyunsaturated fatty acids from phospholipids and glycolipids to triacylglycerols, and an increase in lipid species with 16:0 or 18:1 under nitrogen deprivation conditions. Upregulated stearoyl-ACP desaturase and oleyl-ACP thioesterase promoted the synthesis of 18:1, but restricted acetyl-CoA supply revealed that it was the intensive lipid turnover instead of an attenuated Kennedy pathway that played an important role in the variation in fatty acid composition under early nitrogen deprivation. Finally, two differentially expressed SQUAMOSA promoter-binding proteins (SBPs) were heterologously expressed in Y. lipolytica, demonstrating their role in promoting the accumulation of total fatty acid and the reduction in fatty acid desaturation. These results revealed the crucial role of lipid turnover and SBPs in determining fatty acid desaturation under early nitrogen deprivation, opening new avenues for the metabolic engineering of fatty acid desaturation in microalgae.

Content, composition, and transfer of polyunsaturated fatty acids in an Arctic lake food web

AbstractFreshwater fish production depends on the production and use of polyunsaturated fatty acids (n‐3 and n‐6 PUFA) from lower trophic levels. Here, we aimed to identify the main trophic pathways that support PUFA content in different fish species (mean 39.7 mg/g dry weight) used in the subsistence fishery of the Inuit community in Greiner Lake near Cambridge Bay (Nunavut, Canada). We used stable isotope and taxon‐specific PUFA stocks, to show that the lake food web was divided into distinctive pelagic and littoral benthic food webs and that different fish species obtained their PUFA from different sources within those food webs. The most concentrated fish in n‐3 PUFA was Arctic char (Salvelinus alpinus) that obtained nutritionally valuable PUFA compounds by feeding on pelagic zooplankton rich in the essential fatty acids EPA and DHA and on littoral prey with lower PUFA content. The pelagic consumer, lake whitefish (Coregonus clupeaformis), that fed on mysids and zooplankton was also rich in n‐3 PUFA. The least concentrated in n‐3 PUFA was lake trout (Salvelinus namaycush) that obtained PUFA from low n‐3 PUFA sticklebacks (Pungitius pungitius) and macroinvertebrates and from n‐3 PUFA‐rich littoral mysids. The benthic PUFA were entirely made of n‐6 fatty acids and no n‐3 PUFA were detected. We further quantified that from the mean daily phytoplankton production of 319 mg C·m−2·d−1, 2.9% was assimilated by zooplankton (9.4 mg C·m−2·d−1) and thereby made available to pelagic fish. The food webs to which fish belonged were supported by PUFA produced in the pelagic and benthic zones but likely complemented by inputs from the watershed. The description of the main PUFA pathways of the Greiner Lake food webs explains for the first time the trophic interactions and underlying mechanisms responsible for the health of the fish community in a high‐Arctic lake.

Cross-Ecosystem Linkages: Transfer of Polyunsaturated Fatty Acids From Streams to Riparian Spiders via Emergent Insects

Polyunsaturated fatty acids (PUFAs) are essential resources unequally distributed throughout landscapes. Certain PUFAs, such as eicosapentaenoic acid (EPA), are common in aquatic but scarce in terrestrial ecosystems. In environments with low PUFA availability, meeting nutritional needs requires either adaptations in metabolism to PUFA-poor resources or selective foraging for PUFA-rich resources. Amphibiotic organisms that emerge from aquatic ecosystems represent important resources that can be exploited by predators in adjacent terrestrial habitats. Here, we traced PUFA transfer from streams to terrestrial ecosystems, considering benthic algae as the initial PUFA source, through emergent aquatic insects to riparian spiders. We combined carbon stable isotope and fatty acid analyses to follow food web linkages across the ecosystem boundary and investigated the influence of spider lifestyle (web building vs. ground dwelling), season, and ecosystem degradation on PUFA relations. Our data revealed that riparian spiders consumed considerable amounts of aquatic-derived resources. EPA represented on average 15 % of the total fatty acids in riparian spiders. Season had a strong influence on spider PUFA profiles, with highest EPA contents in spring. Isotope data revealed that web-building spiders contain more aquatic-derived carbon than ground dwelling spiders in spring, although both spider types had similarly high EPA levels. Comparing a natural with an anthropogenically degraded fluvial system revealed higher stearidonic acid (SDA) contents and Σω3/Σω6 ratios in spiders collected along the more natural river in spring but no difference in spider EPA content between systems. PUFA profiles of riparian spiders where distinct from other terrestrial organism and more closely resembled that of emergent aquatic insects (higher Σω3/Σω6 ratio). We show here that the extent to which riparian spiders draw on aquatic PUFA subsidies can vary seasonally and depends on the spider’s lifestyle, highlighting the complexity of aquatic-terrestrial linkages.

Intrauterine Transfer of Polyunsaturated Fatty Acids in Mother-Infant Dyads as Analyzed at Time of Delivery.

Polyunsaturated fatty acids (PUFAs) are essential for fetal development, and intrauterine transfer is the only supply of PUFAs to the fetus. The prevailing theory of gestational nutrient transfer is that certain nutrients (including PUFAs) may have prioritized transport across the placenta. Numerous studies have identified correlations between maternal and infant fatty acid concentrations; however, little is known about what role maternal PUFA status may play in differential intrauterine nutrient transfer. Twenty mother–infant dyads were enrolled at delivery for collection of maternal and umbilical cord blood, and placental tissue samples. Plasma concentrations of PUFAs were assessed using gas chromatography (GC-FID). Intrauterine transfer percentages for each fatty acid were calculated as follows: ((cord blood fatty acid level/maternal blood fatty acid level) × 100). Kruskal–Wallis tests were used to compare transfer percentages between maternal fatty acid tertile groups. A p-value < 0.05 was considered significant. There were statistically significant differences in intrauterine transfer percentages of arachidonic acid (AA) (64% vs. 65% vs. 45%, p = 0.02), eicosapentaenoic acid (EPA) (41% vs. 19% vs. 17%, p = 0.03), and total fatty acids (TFA) (27% vs. 26% vs. 20%, p = 0.05) between maternal plasma fatty acid tertiles. Intrauterine transfer percentages of AA, EPA, and TFA were highest in the lowest tertile of respective maternal fatty acid concentration. These findings may indicate that fatty acid transfer to the fetus is prioritized during gestation even during periods of maternal nutritional inadequacy.

LPIAT1/MBOAT7 contains a catalytic dyad transferring polyunsaturated fatty acids to lysophosphatidylinositol

Human membrane bound O-acyltransferase domain-containing 7 (MBOAT7), also known as lysophosphatidylinositol acyltransferase 1 (LPIAT1), is an enzyme involved in the acyl-chain remodeling of phospholipids via the Lands' cycle. The MBOAT7 rs641738 variant has been associated with the entire spectrum of fatty liver disease (FLD) and neurodevelopmental disorders, but the exact enzymatic activity and the catalytic site of the protein are still unestablished.Human wild type MBOAT7 and three MBOAT7 mutants missing in the putative catalytic residues (N321A, H356A, N321A + H356A) were produced into Pichia pastoris, and purified using Ni-affinity chromatography. The enzymatic activity of MBOAT7 wild type and mutants was assessed measuring the incorporation of radiolabeled fatty acids into lipid acceptors.MBOAT7 preferentially transferred 20:4 and 20:5 polyunsaturated fatty acids (PUFAs) to lysophosphatidylinositol (LPI). On the contrary, MBOAT7 showed weak enzymatic activity for transferring saturated and unsaturated fatty acids, regardless the lipid substrate. Missense mutations in the putative catalytic residues (N321A, H356A, N321A + H356A) result in a loss of O-acyltransferase activity.Thus, MBOAT7 catalyzes the transfer of PUFAs to lipid acceptors. MBOAT7 shows the highest affinity for LPI, and missense mutations at the MBOAT7 putative catalytic dyad inhibit the O-acyltransferase activity of the protein. Our findings support the hypothesis that the association between the MBOAT7 rs641738 variant and the increased risk of NAFLD is mediated by changes in the hepatic phosphatidylinositol acyl-chain remodeling. Taken together, the increased knowledge of the enzymatic activity of MBOAT7 gives insights into the understanding on the basis of FLD.

Acyl-CoA:lysophosphatidylcholine acyltransferases (LPCATs) of Camelina sativa seeds: biochemical properties and function

Main conclusionThe transfer of polyunsaturated fatty acids from phosphatidylcholine to other lipids involves several enzymes. In Camelina sativa seeds, acyl-CoA:lysophosphatidylcholine acyltransferases could be one of the most important players in this process.The transfer of polyunsaturated fatty acids from the location of their synthesis (phosphatidylcholine) to other lipids, e.g., triacylglycerol, remains insufficiently understood. Several enzymes could be involved in this process. One of these enzymes is acyl-CoA:lysophosphatidylcholine acyltransferases (LPCATs). In Camelina sativa seeds, LPCATs could be one of the most important players in this process. Our data clearly indicate that the CsLPCATs present in developing seeds have the potential to transfer almost all polyunsaturated fatty acids synthesised on phosphatidylcholine to the acyl-CoA pool. CsLPCAT activity is the highest at 30 °C, and the enzymes operate well at a pH of 7.0–11.0, with the best activity at a pH of 9.0. The activity of CsLPCATs was inhibited by calcium and magnesium ions at a concentration of 0.05–2 mM. In the forward reaction, CsLPCATs preferentially utilise 18:2-CoA; however, other C18 unsaturated fatty acids are also well accepted. In the backward reactions, there is no clear discrimination between the C18 unsaturated fatty acids utilised by the enzymes for phosphatidylcholine remodelling. The activity of CsLPCATs does not differ much between the stages of seed development.

Study of Total Lipidome of the Sinularia siaesensis Soft Coral

The rapid development of lipidomics of human and higher animals stimulated the study of the lipidome of certain groups of marine organisms. Soft corals are an integral part of the tropical and cold-water ecosystems of the World Ocean, but there is almost no data on the lipidome of these animals. The total lipidome of a tropical soft coral Sinularia siaesensis (Cnidaria: Anthozoa: Octocorallia: Alcionacea) containing intracellular symbiotic microalgae (zooxanthellae) was studied by gas and liquid chromatography-mass spectrometry. The structure and content of 144 molecular species of the main classes of acyl lipids of this alcyonaria were determined, including waxes, triglycerides (TG), monoalkyldiacylglycerides (MADAG), ethanolamine, choline, serine, and inositol glycerophospholipids (PE, PC, PS, and PI), ceramide aminoethylphosphonate (CAEP), sulfoquinovosyldiacylglyceride (SQDG), and mono- and digalactosyldiacylglycerides (MGDG and DGDG). The main components of S. siaesensis lipids were waxes 16:0/16:0, 16:0/18:0, and 18:0/16:0; TG 16:0/16:0/16:0 and 16:2/16:0/16:0; MADAG 18:0e/16:0/16:0, 16:0e/16:0/18:0, and 16:0e/16:0/16:0; and PS 18:0e/24:5, lyso-PC 18:0e, PE 18:1e/20:4, PC 18:0e/20:4, CAEP 18:2b/16:0, SQDG 14:0/16:0, and PI 18:0/24:5. The dominance of saturated waxes in reserve lipids is a species-specific feature of tropical alcyonarians. Waxes are synthesized in the host organism, and zooxanthellae can increase the proportion of saturated waxes by transferring saturated fatty acids (FA). The residues of polyunsaturated FA (PUFA) are found in the molecules of TG and MADAG, mainly in the sn-1 and sn-2 positions, respectively. Detection of MADAG with residues of marker FA of zooxanthellae (18:0e/18:3/16:0, 18:0e/18:4/16:0, and 16:0e/18:3/16:0) confirms the transfer of PUFA from the symbionts to the host. Tetracosapolyenoic FA, which is a chemotaxonomic markers of octocorals, are concentrated in molecular species of PS and PI. Most molecular species of PE, PC, and PS are in alkylacyl form, while for PI molecules, a diacyl form is characteristic. A significant proportion of MGDG, DGDG, and SQDG points to the importance of zooxanthellae in the formation of the lipid profile of symbiotic soft corals. Determination of the profile of lipid molecular species requires the development of a lipidomic approach in the study of biochemistry and ecology of corals and other cnidarians.

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.

Selective transfer of polyunsaturated fatty acids from phytoplankton to planktivorous fish in large boreal lakes

Lake size influences various hydrological parameters, such as water retention time, circulation patterns and thermal stratification that can consequently affect the plankton community composition, benthic–pelagic coupling and the function of aquatic food webs. Although the socio-economical (particularly commercial fisheries) and ecological importance of large lakes has been widely acknowledged, little is known about the availability and trophic transfer of polyunsaturated fatty (PUFA) in large lakes. The objective of this study was to investigate trophic trajectories of PUFA in the pelagic food web (seston, zooplankton, and planktivorous fish) of six large boreal lakes in the Finnish Lake District.Docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and α-linolenic acid (ALA) were the most abundant PUFA in pelagic organisms, particularly in the zooplanktivorous fish. Our results show that PUFA from the n-3 family (PUFAn-3), often associated with marine food webs, are also abundant in large lakes. The proportion of DHA increased from ~4±3% in seston to ~32±6% in vendace (Coregonus albula) and smelt (Osmerus eperlanus), whereas ALA showed the opposite trophic transfer pattern with the highest values observed in seston (~11±2%) and the lowest in the opossum shrimp (Mysis relicta) and fish (~2±1%). The dominance of diatoms and cryptophytes at the base of the food web in the study lakes accounted for the high amount of PUFAn-3 in the planktonic consumers. Furthermore, the abundance of copepods in the large lakes explains the effective transfer of DHA to planktivorous fish. The plankton community composition in these lakes supports a fishery resource (vendace) that is very high nutritional quality (in terms of EPA and DHA contents) to humans.

Seasonal changes in the accumulation of polyunsaturated fatty acids in zooplankton

In aquatic food webs, consumers, such as daphnids and copepods, differ regarding their accumulation of polyunsaturated fatty acids (PUFAs). We tested if the accumulation of PUFAs in a seston size fraction containing different consumers and in Daphnia as a separate consumer is subject to seasonal changes in a large deep lake due to changes in the dietary PUFA supply and specific demands of different consumers. We found that the accumulation of arachidonic acid (ARA) in Daphnia increased from early summer to late summer and autumn. However, ARA requirements of Daphnia appeared to be constant throughout the year, because the accumulation of ARA increased when the dietary ARA supply decreased. In the size fraction .140 mm, we found an increased accumulation of docosahexaenoic acid (DHA) during late summer and autumn. These seasonal changes in DHA accumulation were linked to changes in the proportion of copepods in this size fraction, which may have increasingly accumulated DHA for active overwintering. We show that consumer-specific PUFA demands can result in seasonal changes in PUFA accumulation, which may influence the trophic transfer of PUFAs within the food web.

Accumulation of polyunsaturated fatty acids by cladocerans: effects of taxonomy, temperature and food

Summary1. Zooplankton are important in transferring dietary nutrients, including polyunsaturated fatty acids (PUFA), up through aquatic food webs.2. We tested the hypothesis that the taxonomic composition of zooplankton affects the retention and subsequent transfer of PUFA from upwards through the food web. Using laboratory experiments, we investigated dietary PUFA accumulation and bioconversion capacities of six cladoceran species (Ceriodaphnia sp., Daphnia longispina, Daphnia magna, Daphnia pulex, Scapholeberis mucronata and Simocephalus vetulus) fed on two diets (Scenedesmus obliquus and Cryptomonas sp.) that differed in their PUFA profiles. We performed experiments at two different temperatures (14 and 20 °C) to assess the role of temperature in the trophic transfer of PUFA.3. There was little variation in the concentrations of PUFA in these cladocerans which were controlled by dietary PUFA supply. Moreover, as expected, the concentrations of PUFA in all cladoceran species were higher at low temperature.4. However, even if the composition of PUFA in the cladoceran species generally corresponded to that in their diet, preferential accumulation of some PUFA was recorded in all these taxa. When fed on a highly unsaturated fatty acid‐deficient diet, all the cladocerans showed some ability to convert C18‐PUFA into arachidonic acid and eicosapentaenoic acid. Interspecific variation in the ability to accumulate and bioconvert PUFA in cladocerans was more pronounced at low temperature (14 °C) for both diets.5. Our results strongly suggest that in heterogeneous habitats with food partitioning between co‐existing cladocerans, foraging behaviour may affect the transfer of PUFA more strongly than interspecific variation in accumulating and/or bioconverting dietary PUFA.

Characterization of CoA-independent transacylase activity in U937 cells

Coenzyme A-independent transacylase (CoA-IT) mediates the transfer of polyunsaturated fatty acids from the sn-2 position of a donor phospholipid to the sn-2 position of an acceptor lyso-phospholipid. We have characterized this activity in U937 cells, a human monocytic cell line. The microsomes of these cells contained CoA-IT activity which demonstrated a fatty acid preference for transferring arachidonic acid into exogenously added 1-alkyl-2-lyso-GPC. This enzymatic activity was optimum between pH 6.5 and 9, was heat labile and displayed an apparent Km for 1-alkyl-2-lyso-GPC of 0.4 microM. This activity was not dependent on Ca2+, Mg2+, CoA or ATP, was not inhibited by 2-mercaptoethanol nor by addition of product, 1-alkyl-2-acyl-GPC. The activity of this enzyme was not altered by differentiation of U937 cells towards the macrophage with Me2SO. Treatment of U937 cells with dexamethasone had no effect on transacylase activity. The activity of this enzyme was decreased by the serine esterase inhibitors phenylmethyl-sulfonyl fluoride and N-tosyl-L-phenylalanine chloromethyl ketone and by the histidine modifier diethyl pyrocarbonate, suggesting that CoA-IT may belong to a family of acyltransferase enzymes typified by LCAT. CoA-IT activity was not affected by compounds that affect PLA2 activity, such as quinacrine, aristolochic acid and arachidonic acid, suggesting a mechanism of action for CoA-IT different from classical, low molecular weight PLA2 enzymes. In conclusion, U937 cells contain CoA-IT activity and this study extends our previous knowledge of this enzyme by demonstrating the differences between CoA-IT and PLA2 enzymes and suggesting similarities between CoA-IT and LCAT.

Triacylglycerols and glycerophospholipids in ovaries from maturing and superovulated immature rats.

The concentration and composition of triacylglycerols and glycerophospholipids in the rat ovary were measured during the development of follicles to corpora lutea. Changes occurring during normal development, as the rats aged from 20 to 41 days, were compared with those following treatment of 20-day-old rats with gonadotrophin to stimulate superovulation. In the ovaries of immature superovulated rats, where many follicles developed into corpora lutea in synchrony, the glycerophospholipids increased in concentration throughout growth and luteinization. At the same time the total amount of triacylglycerol rose 10-fold but only increased in concentration during the 24 h after administration of pregnant mare's serum gonadotrophin. In both classes of lipid the proportion of polyunsaturated fatty acids rose throughout. Six days after administration of gonadotrophin 20% of the fatty acid in glycerophospholipid was arachidonic acid (20:4), while the amount of 22:5 acid had risen from 3 to 17% in the triacylglycerols. During normal aging, changes in the developing follicles were masked by the number of other cell types present. In the whole ovary the glycerophospholipids did not change between 20 and 41 days, but the triacylglycerol concentration rose. However, there was a marked similarity between the concentrations of both lipid classes in corpora lutea dissected out of the 41-day-old ovary and those of the superovulated rats. The changes in proportion of the polyunsaturated fatty acids were also the same during normal and stimulated development of the corpora lutea. In the day-41 corpus luteum the phospholipid contained 22% arachidonic acid (20:4), and 20% of the fatty acids in the triacylglycerol was 22:5. The changes in composition of fatty acids suggests that when a follicle begins to develop there is transfer of polyunsaturated fatty acids from cholesterol esters in the interstitial tissue to the triacylglycerols.

Determination of lecithin:cholesterol acyltransfer in mouse plasma and the influence of mercaptoethanol and sulphydryl blocking agents on its activity

1. 1. The cholesterol esterifying activity in mouse plasma has been identified as lecithin:cholesterol acyltransferase (LCAT) on the basis of stoichiometric data, predominant transfer of polyunsaturated fatty acids, wide pH optimum and inhibition of esterification by phospholipase A 2 and sulphydryl blocking agents. The esterifying activity differed from that present in plasma of man, rat and other species since it was partially inhibited by mercaptoethanol and other thiols. 2. 2. Stoichiometric correlations between unesterified cholesterol, lecithin and lysolecithin were not exact, suggesting possible involvement of other enzymes in the overall esterification process during in vitro incubation of mouse plasma. 3. 3. The initial rate of cholesterol esterification was determined by in vitro incubation of mouse plasma, whose cholesterol had been labelled by prior in vivo injection of 3H-mevalonic acid. The mean rate was 281 ± 74 nmol/ml/hr ( mean ± S.D., n = 12 ) and correlated with unesterified cholesterol concentration ( r = 0.73, P < 0.01 ).