Envelope surface ultrastructure and specific gravity of artificially fertilized Pacific cod Gadus macrocephalus eggs

The envelope surface ultrastructure and specific gravity of artificially fertilized eggs of the Pacific cod Gadus macrocephalus were examined. The unfertilized, demersal and slightly adhesive eggs of G. macrocephalus were almost spherical and had no oil globules. Wrinkled envelope surface with elaborated hexagonal reticulated patterns and type I micropyle were observed under a scanning electron microscope. The adhesiveness of the eggs was lost at the blastodermal‐cap stage after fertilization. The micropylar canal was sealed by secretion of the perivitelline fluid, and the entire surface became rough. Numerous bacilli were deposited at the micropyle and the outer envelope surface at the late germ‐ring stage and at the embryo five‐eighths around the yolk stage. The micropyle was completely deformed at the embryo seven‐eighths around the yolk stage. The specific gravity of the fertilized G. macrocephalus eggs ranged from c. 1·0316 to 1·0454. These values, however, sharply decreased towards the end stages of egg development to produce pelagic larvae. The ultrastructural changes in the micropyle and envelope surface of the G. macrocephalus eggs protected the embryo from microorganism infections and mechanical stress during the long incubation period. The adhesiveness and specific gravity of the eggs influenced their dispersion potential.     

Size and visibility of Baltic cod eggs with reference to size-selective and stage dependent predation mortality

Size of Baltic cod eggs from incubation experiments and from field samples was determined by microscopic analysis. Results from plankton samples were compared with corresponding size distributions of cod eggs found in herring stomachs. The influence of fixation on size of different developmental stages was studied. Live eggs from incubation experiments were also sized repeatedly throughout the developmental period with an optical plankton counter (OPC) based on light attenuance measurements as this was assumed to be more closely related to the visibility of the eggs for potential predators than egg diameter as obtained by microscopic analysis.
Preservation in formaldehyde solution caused a small reduction in egg diameter (2.2%) whereby no differences between the developmental stages were detected. Egg size decreased slightly during incubation (6.9%) while the OPC measurements revealed a substantial increase in light attenuance during egg development (42.2%). In the field, a general decrease in egg size with increasing depth was observed while no change between the developmental stages was detectable. The mean size of eggs ingested by herring was slightly lower than in the water column which was most pronounced for the late stages containing a well-developed embryo. The frequency of eggs in an advanced stage of development was considerably higher in the stomachs than in corresponding plankton samples. Therefore, it is suggested that the selection of further developed egg stages by predatory fish in the central Baltic Sea, i.e. herring and sprat, is due to an increase of visibility during egg development in relation to growth and pigmentation of the embryo. Thus it is likely that egg mortality due to predation is stage-dependent rather than strictly size-selective.     

The embryo pancreas is a source of increased yolk amylase in the fertilized eggs of domestic fowls.

The amylases were studied in the yolk of fertilized eggs and in the pancreases of the embryos of domestic fowls. The amylase activity in the yolk increased markedly from 13 days of incubation until hatching, but the activity decreased when the embryos were taken out of the eggs. The isoamylases in the yolk and in the pancreas of the embryo were identical electrophoretically. The amylase occurs mainly in the pancreas of the embryo. We think that the increase in amylase activity in the yolk of fertilized eggs during incubation depends upon the accumulation of pancreatic amylase synthesized by the developing embryo in the egg.     

Characterization of yolk platelets isolated from developing embryos of Arbacia punctulata

Yolk platelets, a major organelle of sea urchin eggs and embryos, were isolated from Arbacia punctulata and biochemically characterized over the course of development to the pluteus stage. Fractionation by sucrose gradient centrifugation revealed yolk platelets in two major density classes. The low-density yolk platelet fraction could be obtained as a very homogeneous preparation and was highly enriched in acid phosphatase activity, while depleted of mitochondrial (cytochrome c oxidase) and plasma membrane (phosphodiesterase) marker enzymes. The chemical composition of low-density yolk platelets prepared from eggs and embryos at various stages of development remained unchanged in terms of phospholipid, triglyceride, hexose, sialic acid, RNA, and protein. However, analysis of the major yolk platelet glycoproteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a number of stage-specific changes. These glycoproteins were found to be major glycoproteins of crude embryo lysates and were predominantly of the polymannose, N-linked type. The predominance of polymannose-type glycoproteins in yolk platelets was further demonstrated by their staining with concanavalin A-colloidal gold in Lowicryl-embedded sections of embryos. These studies represent the first systematic biochemical characterization of intact yolk platelets and the changes in them during early embryonic development.     

Egg yolk triglycerides during development of the domestic chicken

The triglycerides isolated from egg yolk lipids of eggs at various stages of incubation were fractionated according to the degree of unsaturation by argentation chromatography, and individual fractions were analyzed for their fatty acids by gas liquid chromatography. The proportions of the various fractions were constant during development. Fatty acid composition of the fractions were constant also. Fractions with one saturated fatty acid and two monoenoic fatty acids (SM₂) constituted over 40% of the total triglyceride. Palmitic acid constituted over 30%, and oleic acid over 45% of the fatty acid of total triglycerides. It is suggested that during development of thick embryo there is no selective utilization of the egg yolk triglycerides.     

Changes in lipid droplet localization during embryogenesis of the silkworm, Bombyx mori

Lipid droplets are considered one of the most important energy sources in lepidopteran eggs during late embryogenesis, but the process of their incorporation into the embryo is as yet unknown. The present study focused on the process of transition of lipid droplets from the extraembryonic yolk to the embryo of the silkworm Bombyx mori, using morphological and biochemical approaches. The morphological study revealed that the incorporation of lipid droplets from the extraembryonic yolk into the embryo occurs at three points and in three different ways during the development of the embryo. Some lipid droplets were translocated directly from the extraembryonic yolk to the embryo before the blastokinesis stage. However, the majority of lipid droplets together with the other components of the extraembryonic yolk were incorporated in the embryo via both morphogenetic inclusion during dorsal closure and ingestion of the extraembyonic yolk by the developing caterpillar prior to hatching. Similar results were obtained from the biochemical study. Thus, we propose that there are three steps in the incorporation of lipid droplets from the extraembryonic yolk into the embryo. In addition, morphological and biochemical data concerning the total amount of lipid droplets in the egg suggested that lipid droplets were mainly consumed during late embryogenesis, seeming to synchronize with tracheal development.     

Is water uptake by reptilian eggs regulated by physiological processes of embryos or a passive hydraulic response to developmental environments?

Moisture availability is critical for successful embryonic development in many organisms. In most oviparous reptiles, for example, water exchange between eggs and the surrounding environment can have substantial fitness consequences, but regulation of this process is unclear. Here, we evaluate whether water uptake by eggs of the lizard Anolis sagrei is regulated by the presence of a live embryo or is a passive hydraulic response to substrate moisture conditions. Many eggs laid in our captive colony were infertile or contained embryos that died during early stages of development, yet these 'dead' eggs continued to gain mass similar to that of 'live' eggs at least during the first half of incubation. Our results suggest that water uptake by eggs is largely a passive hydraulic process during the first half of incubation, but active regulation by embryos may be necessary during latter stages. Maternal effects (e.g., deposition of salts into yolk) could influence this passive process during early incubation.     

乌骨鸡胚胎发育过程中卵内蛋白质脂肪的变化研究

对乌骨鸡胚胎发育过程中卵内各组成成分(蛋清、蛋黄、胚胎)在不同胚龄(即入孵的第0、3、6、9、12、15、18d和出壳)时蛋白质、脂肪含量变化进行测定与分析．结果表明：在不同胚龄时,蛋清中蛋白质含量显著高于蛋黄,蛋黄中脂肪含量始终高于蛋清;而胚胎中的蛋白质、脂肪逐渐增加。另外,胚胎发育过程中胚胎蛋白质的主要来源是蛋黄中的蛋白质,说明蛋黄是禽类胚胎发育过程中营养需要的主要来源。     

Egg Turning During Incubation has no Effect Upon the Growth of Embryos of Alligator mississippiensis

Alligator eggs are not turned during incubation, instead the embryo adheres to the top inside of the shell. Turning is alleged to shear off the embryo and kill it. Avian egg turning allegedly facilitates embryonic development by stimulating growth of the area vasculosa and minimizing the effects of unstirred yolk and albumen layers. From day 10 to day 45 of incubation, alligator eggs were experimentally turned, gently, through ± 60° in an hourly cycle. This turning regime killed only 6 out of 25 embryos. Compared with unturned controls, no significant effects were observed on the growth, production of extraembryonic fluids or utilization of albumen and yolk for those embryos that survived turning. The protein concentration of amniotic fluid at various stages of alligator development was examined in eggs incubated at 30 and 33°C. The fluid contained very little protein (max <8 mg) at any time: the protein concentration did not change consistently as development progressed. Differences in response to egg turning in birds and reptiles may be associated with the length of the incubation period, the protein content of the albumen and the mechanism of albumen utilization.     

Antioxidants and total oxyradical scavenging capacity during grass shrimp, Palaemonetes pugio, embryogenesis

During embryogenesis in grass shrimp the capacity to scavenge oxyradicals increased as measured by the Total Oxyradical Scavenging Capacity (TOSC) assay. The increase in TOSC during embryogenesis was associated with increasing concentrations of a number of antioxidants, including coenzyme Q (ubiquinone), alpha-tocopherol and reduced glutathione. Glutathione concentrations ranged from 0.004 to 0.005 nmol/embryo in early embryo stages and reached concentrations between 0.16 to 0.23 nmol/embryo in late embryo stages. Ascorbate remained essentially constant (0.16-0.20 nmol/embryo) throughout embryogenesis and may provide the preponderance of TOSC during early embryo development. Carotenoids were associated with yolk lipovitellin and these antioxidants decreased as yolk was absorbed during embryogenesis. Astaxanthin and beta-carotene were identified in embryos with astaxanthin always the principal carotenoid. In early embryo stages there are maternally derived antioxidants but as embryogenesis proceeds there is an assembly of a complex antioxidant system by newly formed cells and tissues.     

Early maternal, genetic and environmental components of antioxidant protection, morphology and immunity of yellow-legged gull (Larus michahellis) chicks

Maternal effects mediated by egg quality are important sources of offspring phenotypic variation and can influence the course of evolutionary processes. Mothers allocate to the eggs diverse antioxidants that protect the embryo from oxidative stress. In the yellow-legged gull (Larus michahellis), yolk antioxidant capacity varied markedly among clutches and declined considerably with egg laying date. Analysis of bioptic yolk samples from clutches that were subsequently partially cross-fostered revealed a positive effect of yolk antioxidant capacity on embryonic development and chick growth, but not on immunity and begging behaviour, while controlling for parentage and common environment effects. Chick plasma antioxidant capacity varied according to rearing environment, after statistically partitioning out maternal influences mediated by egg quality. Thus, the results of this study indicate that egg antioxidants are important mediators of maternal effects also in wild bird populations, especially during the critical early post-hatching phase.     

Ecdysteroid titres and location in developing eggs of Schistocerca gregaria

Ecdysteroid levels in the separated embryo and yolk fractions of Schistocerca gregaria eggs have been determined at each of the developmental stages. The major hormones present both in the free and conjugated state are ecdysone, 20-hydroxyecdysone and 2-deoxyecdysone. At the beginning of embryonic development the ecdysteroids occur only in the yolk whereas, after blastokinesis, they are found in the embryo. The levels of conjugates fall during embryonic development, whereas a decrease of free hormone titres in early embryogenesis is followed by a marked increase in late embryos (stage 26 and 28). The possible role of ecdysteroids in relation to the morphogenetic processes of egg development and the site of origin of the free ecdysteroid peaks are discussed.     

Consequences of prenatal androgen exposure for the reproductive performance of female pheasants (Phasianus colchicus)

Maternal hormones in vertebrate eggs can mediate important forms of maternal effects. However, the function of hormone transfer to the eggs is still debated, especially because long-term fitness consequences have been little studied. We investigated the effect of prenatal exposure to physiologically elevated yolk testosterone (T) levels on reproduction of female pheasants (Phasianus colchicus) in captivity. We found that females hatching from T-injected eggs (T-females) had a lower egg-laying rate than controls, and their eggs were more frequently infertile than those laid by control females. There were no effects of prenatal maternal treatment on egg size and yolk T concentration, but eggs carrying a female embryo laid by T-females had smaller yolks than eggs with a male embryo, while there was no sex difference in yolk size among the eggs laid by control females. Progeny sex ratio was unaffected by maternal treatment. These findings suggest that the transfer of high androgen levels to the eggs by the mother is constrained by complex trade-offs between direct effects on her daughters' reproduction and by trans-generational differential consequences on male and female descendants.     

Embryo lipoproteins and yolk lipovitellin consumption during embryogenesis in Macrobrachium borellii (Crustacea: Palaemonidae)

The prawn Macrobrachium borellii has lecithotrophic eggs with highly-abbreviated development. The major yolk component is lipovitellin (LV), a lipoprotein with 30% lipids (by weight). LV consumption during embryogenesis was followed by ELISA and Western blot analysis using an anti-LV polyclonal antibody. No cross-reacting proteins were observed and LV-like lipoproteins were strongly recognized by the antibody in hemolymph (vitellogenin), yolk (LV) and embryos (LVe), as determined by Western Blot analysis. LV decreased significantly along development from 9.4 to 1.1 microg/mg egg. Consumption rate of LV was slow in early embryogenesis, followed by a rapid utilization in late embryonic stages. Significant LVe amounts were still present at hatching. LV apolipoproteins were selectively degraded during embryo development, being the highest molecular weight subunit the most affected. Comparison among in vitro, in vivo and theoretical proteolysis suggested that trypsin may be involved in LV degradation during late embryogenesis. Embryo lipoprotein (HDLe) synthesis was first detected at stage 6. HDLe shared the same density, MW and subunit composition as adult hemolymph HDL(1) and did not cross-react with LV-like lipoproteins. Though expressed at low concentration, it fulfilled embryo needs for lipid transport among organs.     

Magnetic resonance imaging study of the structure of the yolk in the developing avian egg

Magnetic resonance imaging (MRI) techniques were used to study the morphology of the latebra and concentric rings seen in the yolk of White Leghorn eggs during development of the avian embryo. Previous studies of the macroscopic structure of avian yolk have revealed the latebra, a vase-shaped structure beneath the blastoderm composed of white yolk. The bulbous portion in the center of the yolk is termed the body of the latebra. The thinner portion extending toward the blastoderm is referred to as the neck of the latebra. As the neck of the latebra approaches the blastoderm, it flares out to become the nucleus of Pander. The remainder of the yolk often features alternating concentric layers of white and yellow yolk. These layers, which appear as rings in sections, are thought to represent the daily accumulation of yolk during oogenesis. In this study eggs were imaged with a single slice spin echo sequence using MRI parameters that maximized the visualization of the latebra and concentric rings in the egg yolk. Some experiments were conducted for 2 to 3 day periods with eggs kept in the bore of the magnet using a small incubator that was constructed using a temperature-controlled water pump. The concentric rings of the yolk and the body of the latebra flatten and become more elliptical during development. The neck of the latebra becomes shorter and disappears around the 7th day of incubation. The body of the latebra starts to become incorporated into the embryo at about the 7th day of incubation and usually disappears by the 13th day. The concentric rings are no longer visible as distinct entities at this time. Histochemical procedures carried out as a result of MRI findings indicate that the latebra is an iron-rich structure.     

Developmental fate of the yolk protein lipovitellin in embryos and larvae of winter flounder, Pleuronectes americanus.

The developmental fate of the vitellogenin-derived yolk protein, lipovitellin (Lv), was investigated in winter flounder embryos and yolk-sac larvae. Since Lv is present as only one major polypeptide in ovulated winter flounder eggs, unlike the multiple yolk polypeptides found in the mature eggs of most teleosts, this system is presented as a simpler model of yolk protein structure and utilization during teleostean development. Winter flounder Lv is cleaved during embryogenesis from a 94 kD polypeptide at fertilization to 67 kD and 26 kD polypeptides at hatching. The rate of this proteolytic processing is slow during early embryonic development, but enters a more rapid phase between days 8 and 12 post-fertilization in embryos reared at 4-5 degrees C, and approaches 50% completion at day 10. Lv processing is essentially complete 3 days before hatching; nevertheless, major degradation of the Lv peptide by the developing winter flounder does not occur until after hatching. The Stokes radius of Lv changes only moderately following processing, from 4.50 nm in unfertilized eggs to 4.19 nm in late embryos and newly hatched larvae, whereas the processed Lv retains its heat stability relative to other yolk polypeptides. Nearly 50% of its lipid content, however, is released from the Lv particle during embryogenesis, concomitant with cleavage of the Lv 94 kD polypeptide. Lv processing may thus render a portion of the yolk protein-associated lipid more accessible to the developing embryo, whereas other yolk components are retained for later use by the winter flounder larva. Alternately, removal of lipid may lead to proteolytic vulnerability of the Lv polypeptide. In either case, only a portion of the lipid moiety of the Lv particle appears to play a significant nutritive role for the embryo, whereas its protein component is reserved for larval use. J. Exp. Zool. 284:686-695, 1999.     

Organisation of Xenopus egg cytoplasm: response to simulated microgravity

The cytoplasm of Xenopus fertilised eggs appears to be organised into three major compartments based primarily on the uneven distribution of yolk platelets. There is a shift of these yolk compartments during the first cell cycle that is thought to be involved in the dorsal/ventral morphogenesis of the embryo. The involvement of gravity in Xenopus cytoplasmic organisation and in compartment shifts was addressed by examining, cytologically, the yolk compartments in embryos that developed under the simulated microgravity conditions of the horizontal clinostat. The cytoplasmic organisation into yolk compartments was found to be maintained, and the asymmetric movements of compartments still occurred in eggs that developed on the clinostat. It is suggested that the organisation of Xenopus egg cytoplasm into discrete compartments relies on forces other than those involving gravity (i.e., not density differences), and that the compartment shifts that take place during the first cell cycle are active movements. The variation in compartment size and composition observed from batch to batch of eggs, and to a lesser extent from egg to egg, during this study was addressed.     

Thyroid Hormone Deposition in Avian Eggs and Effects on Embryonic Development

Studies to date indicate that thyroid hormones are present inthe eggs of chickens and quail and that those hormones are primarilyin the yolk. Quail hens deposit thyroid hormones into eggs inproportion to their own thyroid status, but appear to show someregulation of this process. Indirect studies suggest that thyroidhormones are transferred into oocytes bound to lipoproteinsand transthyretin, both of which are taken up by receptor-mediatedprocesses. Thyroid hormones bound to yolk lipoproteins may enterembryos with yolk that is taken up by non-specific endocytosisor they may be transported into the embryo by specific carriers.To date most of these ideas about thyroid hormone transportinto egg yolk and from egg yolk into embryos have not been investigateddirectly. In quail, very high T₄ content of eggs is associatedwith accelerated differentiation and growth of embryonic pelviccartilage, a thyroid hormone-responsive tissue. We evaluatethese effects on embryonic tissues and the changes in yolk hormonecontent during incubation in relation to the timing of thyroiddevelopment and studies of the capability for tissue responsesto thyroid hormones during early embryonic life     

Embryonic growth and mobilization of energy and material in oviposited eggs of the red-necked keelback snake, Rhabdophis tigrinus lateralis

We used the red-necked keelback (Rhabdophis tigrinus lateralis) as a model animal to study embryonic growth and mobilization of energy and material in oviposited snake eggs. Females (N=12) laid eggs between late May and early June. Eggs were incubated at 30 (+/-0.3) degrees C. One egg from each clutch was dissected at five-day intervals starting at oviposition. Incubation length averaged 27.9 days. Three phases of embryonic growth or yolk depletion could be detected in this study. The first phase, between oviposition and Day 10, was one of minimal transfer of energy and material from yolk to embryo. The second phase, between Day 10 and Day 22-23, was characterized by increasingly rapid embryonic growth and yolk depletion. The third phase, between Day 22-23 and hatching, was characterized by a gradual reduction in embryonic growth and yolk depletion. Approximately 73.6% of dry mass, 50.0% of non-polar lipids and 57.8% of energy were transferred from egg to embryo during incubation. Embryos withdrew mineral from the eggshell mainly during the last quarter of incubation. Our data show that oviposition does not coincide with the onset of rapid embryonic growth in oviparous species of squamate reptiles that are positioned midway within the oviparity-viviparity continuum, and that the greater conversion efficiencies of energy and material from egg to hatchling in snakes can be mainly attributed to their lower energetic costs of embryonic development and greater residual yolk sizes.