Brian K. Speake

Tissue-specific differences in antioxidant distribution and susceptibility to lipid peroxidation during development of the chick embryo

The purpose of this study was to determine the tissue-specific acquisition of antioxidant capacity during chick embryo development and to assess the effectiveness of this process in the prevention of lipid peroxidation. The transfer of alpha-tocopherol, carotenoids and ascorbic acid from the yolk/yolk sac membrane (YSM) to the developing chick embryo and the distribution of these antioxidant compounds between the embryonic tissues were investigated. The concentrations of alpha-tocopherol and carotenoids in the yolk decreased between day 15 of development and hatching at day 21, concomitant with an increase in the levels of these antioxidants in the YSM. The concentration of both these lipid-soluble antioxidants in the liver increased dramatically between day 18 of embryonic development and day 1 after hatching. The adipose tissue content of alpha-tocopherol also increased markedly during the late embryonic/early neonatal period. However, the levels of alpha-tocopherol in the liver were far higher than in any other tissue with particularly low levels observed for the brain. Also, carotenoids were undetectable in the developing brain. Ascorbic acid was not present in the initial yolk but high levels of this water-soluble antioxidant were detected in the YSM, particularly at the early stages of development. The concentration of ascorbic acid in the embryonic brain was far higher than in any other tissue. Homogenates of brain tissue were extremely susceptible to lipid peroxidation during incubation in vitro whereas extracts of liver, yolk and YSM were relatively resistant to lipid peroxidation, particularly in the absence of exogenous Fe2+.

Distribution of carotenoids from the yolk to the tissues of the chick embryo

Abstract Laying hens were placed on either a control diet or a carotenoid-supplemented diet. The subsequent concentrations of carotenoids in the yolks of the newly laid fertile eggs (μg carotenoid/g fresh yolk) were 13.3 and 41.1 on the control diet and high-carotenoid diet, respectively. For the day-old chicks derived from the high-carotenoid group, the carotenoid levels in the yolk sac membrane, liver, and plasma were approximately 3 times greater than in the control group; however, the other tissues were less responsive. The proportion of lutein (% wt/wt of total carotenoids) was far lower in the liver of the chick than in the yolk. However, in the non-hepatic tissues the proportion of lutein was generally higher than in the yolk. This suggests that discrimination between different carotenoids may occur in the embryo. For the high-carotenoid group, the peroxidative susceptibility of extracts of the yolk, yolk sac membrane, and liver was decreased compared with samples from the control group. Thus, carotenoids may provide antioxidant protection to the tissues.

Designer egg evaluation in a controlled trial

Objective: To evaluate the ability of designer eggs enriched in vitamin E, lutein, selenium (Se) and docosahexaenoic acid (DHA) to deliver micronutrients to the human in a palatable and visually acceptable form.Design: Double-blind, placebo-controlled trial, two treatment groups balanced for sex and age.Setting: Department of Biochemistry and Nutrition, SAC, Scotland.Subjects: Forty healthy adult volunteers completed the study. Volunteers were recruited among staff of the Scottish Agricultural CollegeInterventions: Volunteers consumed, for 8 weeks, either a designer egg or a normal table egg per day. Fasting blood samples were taken before and at the end of the study.Results: Consumption of designer eggs enriched in vitamin E, lutein, Se and DHA significantly increased the levels of α-tocopherol, lutein and DHA in plasma as compared to the changes found after consumption of normal table eggs, with the largest increases found in plasma lutein (1.88-fold increase). The proportion of DHA was increased in all the main lipid classes of the plasma including triacylglycerol (2.3-fold), free fatty acids (1.6-fold), cholesteryl ester (1.4-fold) and phospholipid (1.3-fold). Egg consumption did not change Se concentration in plasma, blood pressure, total plasma lipid concentrations or the concentrations of total cholesterol and HDL-cholesterol in plasma.Conclusion: Consumption of designer eggs enriched in vitamin E, lutein, DHA and Se as part of normal diet for 8 weeks effectively increased the blood levels of α-tocopherol, lutein and DHA.Sponsorship: Scottish Office Agriculture, Environment, and Fisheries Department.European Journal of Clinical Nutrition (2000) 54, 298–305

Comparison of nutrient transport across the placenta of lizards differing in placental complexity.

We have reviewed published and new quantitative data on the net uptake of nutrients by embryos of oviparous and viviparous lizards that vary in chorioallantoic placental complexity to better understand the evolution of complex placentae. We assessed net nutrient uptake during embryonic development by measuring the total dry mass, or the mass of separate nutrients, in the egg at about the time of ovulation and in the neonate. There is no significant difference in the fresh egg to neonate dry mass ratio of oviparous and viviparous species that have simple placentae, indicating that there is little, if any, net uptake of nutrients by viviparous species with simple chorioallantoic placentae. In contrast, there is significant uptake of dry matter and individual nutrients across the placenta of species with complex chorioallantoic placentae. Species of the genus Niveoscincus have a range of placentae and nutrient uptakes, even among populations of one species, suggesting that further studies among populations of single species are required. Data are available for relatively few clades, and all the data for the three most complex chorioallantoic placental types are derived from a single genus. Thus, further research on new genera of lizards is required to overcome the potentially confounding effects of phylogeny in our analyses.

Carotenoid discrimination by the avian embryo: a lesson from wild birds

The concentrations (microg/g wet yolk) of total carotenoids in eggs of the common moorhen (Gallinula chloropus), American coot (Fulica americana) and lesser black-backed gull (Larus fuscus), collected in the wild, were 47.5, 131.0 and 71.6, respectively. In contrast to data for eggs of the domestic chicken, beta-carotene was a significant component in the yolks of these three wild species, forming 25-29% by wt. of the total carotenoids present. The concentration of total carotenoids in the livers of the newly-hatched chicks was 5-10 times higher than in the other tissues and beta-carotene was again a major component, forming 37-58% of the hepatic carotenoids. In the newly-hatched gull, the proportions of both lutein and zeaxanthin were very low in the liver but high in the heart and muscle when compared with the yolk. By contrast canthaxanthin, echinenone and beta-carotene were very minor constituents of heart and muscle when compared with their proportions in the yolk of the gull. The proportions of lutein and zeaxanthin in the liver of the newly-hatched coot and moorhen were also far lower than in the yolk whereas the liver was relatively enriched with beta-cryptoxanthin, beta-carotene and (in the moorhen) echinenone. The results indicate that avian embryos discriminate between different carotenoids during their distribution from the yolk to the various tissues.

Maternal docosahexaenoic acid supplementation and fetal accretion

Docosahexaenoic acid (DHA) (22 : 6n-3) is a polyunsaturated fatty acid that is an essential constituent of membranes, particularly of the nervous system. Infants acquire DHA from their mothers, either prenatally via the placenta or postnatally in milk. The present study aimed to test the hypothesis that maternal supplementation during the second and third trimesters of pregnancy enriches maternal and/or fetal DHA status. In a randomised, prospective, double-blind study 100 mothers received either fish-oil capsules containing 400 mg DHA/g (200 mg/d) (n 50), or placebo containing 810 mg oleic acid/g (400 mg/d) (n 50) from 15 weeks gestation until term. Venous blood samples were obtained from mothers at 15, 28 and 40 weeks, and from the umbilical cord at birth. Total fatty acids in plasma and erythrocytes were analysed by GC-MS. There were no significant differences between maternal groups in baseline DHA, as a proportion of total fatty acids (g/100 g total fatty acids) or concentration (nmol/ml), in plasma and erythrocytes. DHA concentrations in plasma at 28 weeks (P=0.02) and erythrocytes at both 28 weeks (P=0.03) and term (P=0.02) were 20 % higher in supplemented mothers than the placebo group. DHA accounted for a higher proportion of total fatty acids in erythrocytes of supplemented mothers at 28 weeks (P=0.003) and term (P=0.01). There were no significant differences between groups in DHA (g/100 g total fatty acids or nmol/l) in cord blood. Maternal DHA status was maximal in mid-trimester and declined to term, at a lower rate in supplemented compared with unsupplemented mothers. Maternal DHA supplementation significantly increases maternal DHA status and limits the last trimester decline in maternal status, aiding preferential transfer of DHA from mother to fetus.

A review of the evolution of viviparity in lizards: structure, function and physiology of the placenta.

The aim of this review is to collate data relevant to understanding the evolution of viviparity in general, and complex placentae in particular. The wide range of reproductive modes exhibited by lizards provides a solid model system for investigating the evolution of viviparity. Within the lizards are oviparous species, viviparous species that have a very simple placenta and little nutrient uptake from the mother during pregnancy (lecithotrophic viviparity), through a range of species that have intermediate placental complexities and placental nutrient provision, to species that lay microlecithal eggs and most nutrients are provided across the placenta during development (obligate placentotrophy). In its commonest form, lecithotrophic viviparity, some uptake of water, inorganic ions and oxygen occurs from the mother to the embryo during pregnancy. In contrast, the evolution of complex placentae is rare, but has evolved at least five times. Where there is still predominantly a reliance on egg yolk, the omphaloplacenta seems to be paramount in the provision of nutrition to the embryo via histotrophy, whereas the chorioallantoic placenta is more likely involved in gas exchange. Reliance on provision of substantial organic nutrient is correlated with the regional specialisation of the chorioallantoic placenta to form a placentome for nutrient uptake, particularly lipids, and the further development of the gas exchange capabilities of the other parts of the chorioallantois.

Effect of Vitamin E and Selenium Supplementation of Cockerel Diets on Glutathione Peroxidase Activity and Lipid Peroxidation Susceptibility in Sperm, Testes, and Liver

The phospholipids of avian spermatozoa are characterized by high proportions of arachidonic (20:4n-6) and docosatetraenoic (22:4n-6) fatty acids and are therefore sensitive to lipid peroxidation. α-Tocopherol and glutathione peroxidase [GSH-Px] are believed to be the primary components of the antioxidant system of the spermatozoa. The present study evaluates the effect of vitamin E and vitamin E plus Se supplementation of the cockerel diet on GSH-Px activity, vitamin E accumulation, and lipid peroxidation in the spermatozoa, testes, and liver. At the beginning of the experiment 75 Rhode Island Red cockerels were divided into five groups, kept in individual cages, and fed a wheat-barley-based ration balanced in all nutrients. Supplements fed to the different groups were as follows: vitamin E, 0, 20, 200, 20, and 200 mg/kg to groups 1–5, respectively, with groups 4 and 5 also receiving 0. 3 mg Se/kg. The vitamin E supplementation produced increased levels of α-tocopherol in semen, testes, and liver. The inclusion of the Se into the cock diet had a significant (P < 0.01) stimulating effect on GSH-Px activity in seminal plasma, spermatozoa, testes, and liver. The increased vitamin E concentration in the spermatozoa was associated with a reduction in their susceptibility to lipid peroxidation. Similarly, the increased GSH-Px activity provided enhanced protection against lipid peroxidation.

Relationship between vitamin E content and susceptibility to lipid peroxidation in tissues of the newly hatched chick

1. The effect of supplementing the diet of the parent hen with vitamin E on the vitamin E content of the yolk and of embryonic and neonatal tissues was evaluated and the effects of elevated tissue concentrations of vitamin E on peroxidation susceptibility was examined. 2. Laying hens (Ross 1 broiler-breeder strain) were maintained on diets containing either 147 (control diet) or 365 (high vitamin E diet) microg vitamin E/g feed. 3. In the day-16 embryo, the concentrations of of vitamin E in the yolk sac membrane, liver, brain and lung were respectively 5.0, 4.3, 1.7 and 5.6 times greater for those derived from the hens on the high vitamin E diet compared with those from the control group. 4. In the day-old chick, the concentrations of vitamin E in the yolk sac membrane, liver, brain and lung were respectively 14.8, 2.8, 3.0 and 5.1 times greater for those derived from hens on the high vitamin E diet compared with those from the control group. 5. Homogenates of tissues from the day-old chick were incubated in the absence and presence of Fe2+ in order to determine the extent of spontaneous and iron-stimulated peroxidation as measured by the generation of thiobarbituric acid reacting substances. For the chicks derived from hens on the control diet, the brain was markedly more susceptible to both spontaneous and iron-stimulated peroxidation than were the other tissues. Tissues from the chicks derived from the hens on the high vitamin E diet exhibited significantly reduced susceptibilities to peroxidation. In particular, the susceptibility of the brain was reduced to the same level as that of the other tissues. 6. It is concluded that the high peroxidative susceptibility of the chicks brain can be normalised by supplementation of the parent hen with vitamin E.

Tissue-specific antioxidant profiles and susceptibility to lipid peroxidation of the newly hatched chick.

The hatching process is characterized by a range of adaptive changes, and a newly hatched chick is considered as an intermediate stage between prenatal and postnatal development. The aim of the present study was to evaluate the characteristic relationships between tissue-specific fatty acid composition and antioxidant protection in newly hatched chicks. Liver, yolk sac membrane, heart, kidney, lung, and four brain regions (cerebrum, cerebellum, stem, and optic lobes) were collected. Fatty acid composition of total lipids and phosphoglycerides, α-tocopherol, lutein, ascorbic acid, reduced glutathione, and the activities of Mn-and Cu,Zn-superoxide dismutase (SOD) and Se-dependent and non-Se-glutathione peroxidase (GSH-Px), and catalase (CAT) were determined. The levels of Fe, Cu, Zn, and Mn as well as tissue susceptibility to lipid peroxidation were also studied. The tissues of the newly hatched chick showed distinctive features in fatty acid profiles, antioxidant accumulation, and susceptibility to lipid peroxidation. The brain clearly displayed the greatest susceptibility to spontaneous and Fe-stimulated lipid peroxidation, was highly unsaturated and contained very low levels of vitamin E, no detectable carotenoids, low GSH-Px, and low CAT activity. At the same time, the brain was characterized by high ascorbic acid concentration and comparatively high SOD activity. It was suggested that in postnatal development, antioxidant enzymes presumably play the major role in antioxidant protection of the chick tissues.