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PubMed1983

Influence of egg investments on in-vitro penetration of mouse eggs by misshapen spermatozoa.

Three groups of eggs, intact, cumulus-free and zona-free, were inseminated in vitro with spermatozoa from C57BL/Kw male mice and incubated for 1-2 h. In fixed, toluidine blue-stained preparations, spermatozoa surrounding the eggs or attached to them remained colourless, while sperm heads that had penetrated the vitellus were deeply stained. Those in the early stage with still condensed chromatin were used to evaluate the head shapes. In zona-free and in cumulus-free eggs, only severely misshapen sperm heads appeared at a lower frequency than in the samples used for insemination. The proportion of abnormal sperm heads of all classes was significantly lower in intact than in cumulus-free eggs and most severely abnormal forms were not found in intact eggs. The results suggest that abnormal spermatozoa are able to participate in fertilization, although their chances of penetrating the ova in vitro are greatly reduced in the presence of egg investments, especially of the cumulus.

Age Factors↗

PubMed1992

Ability of thawed tiger (Panthera tigris) spermatozoa to fertilize conspecific eggs and bind and penetrate domestic cat eggs in vitro.

Electroejaculates from tigers were collected and half was used fresh to inseminate tiger eggs in vitro and domestic cat eggs stored in a hypertonic salt solution. The remainder was pelleted, frozen in a solution of 20% egg yolk, 11% lactose and 4% glycerol, thawed and cultured with tiger and domestic cat eggs. The motility index ((sperm % motility)+(status rating x 20))/2 for thawed spermatozoa was about 86% of that in fresh aliquots. Of the 49 tiger oocytes inseminated in vitro with fresh spermatozoa, 34 (69.4%) cleaved, compared with 33 of 47 oocytes (70.2%) cultured with thawed spermatozoa (P > 0.05). Embryos generated by either sperm treatment could develop in vitro to the 16-cell or morula stage. Fresh and thawed tiger spermatozoa were equally capable (P > 0.05) of binding and penetrating the outer and inner zona pellucida of domestic cat eggs. These results demonstrate the ability of frozen-thawed tiger spermatozoa to (i) penetrate homologous and heterologous eggs and (ii) result in conspecific, advanced development of preimplantation embryos in vitro.

Animals↗

PubMedDate not supplied

Microinjection of human sperm into perivitelline space of hamster eggs: comparison with zona-free hamster egg penetration of human sperm.

Micromanipulation of human sperm and oocyte has been utilized to facilitate fertilization of those patients with male factor due to oligoasthenospermia or those patients with repeated fertilization failure in an in vitro fertilization (IVF) program. Before manipulating human gametes, one needs experience with animal models. Our objective was to perform subzonal insertion of human sperm into hamster eggs and to compare the result with that of sperm penetration assay (SPA) using zona-free hamster eggs. Semen samples were obtained from 15 fertile donors with normal semen analysis and the motile sperm were collected by swim-up procedure. Microinjection was performed by injecting a varied number of sperm into the perivitelline space of 222 hamster eggs pretreated with sucrose solution (0.1 M). The rate of damage of eggs during microinjection was 7.2% (16/222). The rates of penetration in the microinjection group were 5.1% (4/79) for 1-5 sperm injected, 10.9% (11/101) for 6-10 sperm injected, and 11.5% (3/26) for 11-15 sperm injected. The average rate of penetration per egg was 8.7% (18/206), and the polyspermic rate was 11.1% (2/18). Simultaneously SPA was performed in each sample of semen as a positive control, and the average rate of penetration of SPA was 51.4% (108/210). The rate of penetration in the microinjection group was significantly smaller (p < .05) than that in the SPA group. Whether the penetration rate and polyspermic rate in a hamster model reflect similar results in human oocyte requires further investigation. However, the hamster egg provides an ideal model to develop a micromanipulation technique for human beings.

Animals↗

PubMed1987

Changes in ring-necked pheasants' (Phasianus colchicus) egg formation time, oviposition lag time, and egg sequence length due to ahemeral light-dark cycles.

A total of 108 pheasant hens was exposed to either a conventional 24-hr (14L:10D), an ahemeral 22-hr (14L:8D), or an ahemeral 26-hr (14L:12D) light-dark (L:D) cycle. Total lag time for each egg sequence was greater in the ahemeral cycles than in the conventional L:D cycle, resulting in significant (P less than .05) differences in egg formation times (EFT) or intraclutch intervals of 26.4, 25.8, and 27.5 hr, respectively. The EFT under the ahemeral 26-hr L:D cycle was synchronized (within 1.5 hr) with the length of the L:D cycle. This light cycle resulted in the longest egg sequences (average 8.5 eggs per sequence) produced under any of the three L:D cycle treatments due to a shorter pause in oviposition. Values were significantly (P less than .05) different from the average values obtained under the conventional 24 hr L:D cycle (4.9 egg per sequence) and ahemeral 22 hr L:D cycle (3.5 eggs per sequence).

Animals↗

PubMed1991

Two-way selection for threshold body weight at first egg in broiler strain females. 2. Effect of supplemental light on weight and age at first egg.

In order to undergo the processes leading to lay, female chickens must reach a minimum threshold age and body weight, and be exposed to some minimum amount of light stimulation. By two-way selection over six generations lines were developed that differed markedly in body weight at first egg. The effect of supplemental light on age and body weight at first egg of these lines was studied in two experiments. In the first experiment, birds of the two lines were provided ad libitum access to feed from hatch, reaching the weight threshold at an early age. Supplemental light was provided at either 91 or at 126 days of age (before or after the age threshold, respectively). Differences in body weight and age at first egg of the two lines were the same in both light treatments. In the second experiment, birds of the two lines were raised under feed restriction to 154 days of age and then released to ad libitum feeding under supplemental or natural (autumn) light. Differences between the lines in age and weight at first egg were greater in the group maintained under natural light than under supplemental light. The results are interpreted in terms of a model relating age at first egg to weight and age thresholds for onset of competence to respond to light stimulation, and to cumulative light stimulation required from onset of competence to first egg. It is proposed that selection affected the total amount of light stimulation required from onset of competence to onset of lay, but did not affect weight or age thresholds for onset of competence.

Aging↗

PubMed1992

Thapsigargin activates a calcium influx pathway in the unfertilized mouse egg and suppresses repetitive calcium transients in the fertilized egg.

At fertilization, the sperm initiates development of the mouse egg by inducing a large transient increase in the intracellular Ca2+ concentration ([Ca2+]i), which is followed by repetitive transient increases in [Ca2+]i. To determine how the repetitive Ca2+ transients are produced, thapsigargin, an inhibitor of the endoplasmic reticulum Ca-ATPase, was used to deplete intracellular Ca2+ stores within the egg. In the unfertilized egg, thapsigargin (1-50 microM) caused a slowly rising and falling transient increase in [Ca2+]i with or without extracellular Ca2+. An influx pathway for Ca2+ is activated by thapsigargin, since an immediate increase in [Ca2+]i occurred when Ca2+ was added to eggs after thapsigargin treatment in a Ca2+, Mg(2+)-free medium. This suggests that Ca2+ entry in the mouse egg may be coupled to the emptying of an intracellular store. The magnitude of the first Ca2+ transient at fertilization was reduced by as much as 84% in eggs pretreated with thapsigargin. Reduction of extracellular Ca2+, by addition of a Ca2+ chelator, suppressed the repetitive Ca2+ transients following fertilization. The Ca2+ transients also require filling of an intracellular store; they were suppressed when thapsigargin was added before or after fertilization. These results support the hypothesis that the first sperm-induced Ca2+ transient at fertilization depletes an intracellular Ca2+ store, triggering an increase in plasma membrane Ca2+ permeability, and that the enhanced Ca2+ influx causes repetitive Ca2+ transients due to the periodic filling and emptying of an intracellular Ca2+ store.

Adenosine Triphosphatases↗

PubMed1992

Post-fertilization changes in Discoglossus pictus (Anura) eggs result in the formation of a capsular chamber where the egg rotates.

Discoglossus pictus is one of the few anurans with an egg where a capsular chamber forms as a consequence of fertilization; the egg with its vitelline envelope rotates in this chamber according to gravity. We investigated the formation of the capsular chamber through various experimental cytochemical and ultrastructural approaches, and found that it is the product of plug liquefaction. The plug is a lens-shaped jelly coat typical of Discoglossus, and covering only part of the egg animal half. About 15 min after fertilization, granular material coming from the egg enters the plug, which gradually dissolves and, once liquefied, reorganizes itself around the entire egg, thus forming the chamber. This process goes through stages of rearrangement of the 25-A- and 250-A-thick filaments which constitute the plug matrix. The material entering the plug derives from the exocytosis of two vacuole types, with electron transparent and granular PAS-positive contents. Liquefaction of the plug correlates with the reduction of disulfide bonds present in its matrix. Furthermore, in vitro tests showed that the substances released from the egg are active in selectively dissolving only the plug, and lose activity upon boiling.

Animals↗

PubMedDate not supplied

[The functional characteristics of the ultrastructure of the outer membranes of opisthorchid eggs. 2. The mechanisms of egg retention on substrates].

The studies of the functional features of the ultrastructure of the outer membranes of Opisthorchis felineus and Clonorchis sinensis eggs have been continued. The variants of egg behavior on different substrates (artificial, natural, organic and inorganic) have been experimentally studied. It has been established that O. felineus and C. sinensis eggs retain on a number of substrates due to covering with microflora (e.g. fungus mycelium clinging to the rubbed surface of the egg outer membranes); sticking in the uneven surface of the substrate; catching in the uneven surface of the substrate; catching on microfibers of plant fragments. The features found are to a great extent caused by a complex comb-like structure of egg membranes and are interpreted in terms of the earlier unknown mechanisms of parasite adaptation (at the egg stage) facilitating its contact with molluscs--their intermediate hosts.

Adhesiveness↗

PubMed1991

Application of alcohol- or acetone-fixed Schistosoma eggs as an alternative to lyophilized eggs for the circumoval precipitin test.

Antigenicity of Schistosoma mansoni and S. japonicum eggs preserved in ethanol or acetone were assessed in a circumoval precipitin (COP) assay. The egg antigens were found to retain sufficiently their COP reactivity for the diagnosis of both schistosomiasis mansoni and japonica, although their reactivity became lower than that of lyophilized eggs. These alternative preparations for COP tests have advantages, such as keeping eggs directly in fixatives soon after the egg-purification process. Furthermore, evaporation-process may cause eggshell cleavages which facilitate the reaction. The possible usefulness of those eggs in COP assays in local endemic areas is discussed.

Acetone↗

PubMedDate not supplied

Effect of Mycoplasma gallisepticum bacterin on egg-transmission and egg production.

Groups of white leghorn hens were vaccinated twice with a Mycoplasma gallisepticum (MG) bacterin, once with bacterin, or left unvaccinated. Four weeks after vaccination, they were challenged with virulent R strain MG. Egg production was significantly higher in challenged vaccinated groups than in the challenged control group. Four challenged control hens went out of production, whereas only one twice-vaccinated hen did. MG was first isolated directly from eggs 5 days postchallenge (PC) in twice-vaccinated hens, 4 days PC in once-vaccinated hens, and 2 days PC in controls, and the hens continued to lay positive eggs till the end of the experiment 7 weeks PC. MG was found in 17.65%, 38.55%, and 45.90% of eggs cultured in twice-vaccinated, once-vaccinated, and control groups, respectively. Nine of 16 twice-vaccinated hens were found to be shedding MG through their eggs, whereas 15 of 17 once-vaccinated hens and 14 of 16 controls were shedding MG through their eggs.

Animals↗

PubMedDate not supplied

Mycoplasma gallisepticum vaccination: further studies on egg transmission and egg production.

Leghorn hens vaccinated twice with an inactivated Mycoplasma gallisepticum (MG) bacterin before egg production and subsequently challenged with virulent MG were protected against transmission of MG through the egg. Unvaccinated control hens transmitted MG through the egg at a high rate. When unvaccinated hens were vaccinated with MG bacterin 2 weeks after challenge with MG, there was no significant decrease in egg transmission. Hens vaccinated twice before laying did not suffer as severe egg-production drops as unvaccinated hens did when challenged with virulent MG. In a natural MG outbreak in a leghorn breeder flock, 50 hens were separated from the remainder of the flock and vaccinated with MG bacterin approximately 3 weeks after initial exposure. The unvaccinated hens transmitted MG through the egg at a rate three times higher than the rate of transmission of the vaccinated hens.

Animals↗

PubMed1995

Biological activity assays of cell-free reassembled nuclei--injecting cell-free reassembled nuclei into unfertilized eggs can induce the eggs to cleave and reconstitute asters, and the injected nuclei undergo cell cycle changes.

Nucleus may reassemble spontaneously in cell-free mixture of HeLa metaphase chromosomes, Xenopus egg extracts and ATP-regenerating system, and the nucleus shows some biological activities. It is found that, after being injected into unfertilized mature eggs, the cell-free reassembled nuclei can cause the eggs to cleave and reconstitute asters in their cytoplasm, and the injected nuclei undergo changes in response to cell cycle regulators stored in the eggs, and that reinjecting cytostatic factors (CSF) into the eggs can stabilize the eggs in mitotic phase, cause the nuclei disassembly and chromatin condensation to chromosomes.

Animals↗

PubMed1996

Recombinant IL-10 and IL-10/Fc treatment down-regulate egg antigen-specific delayed hypersensitivity reactions and egg granuloma formation in schistosomiasis.

In infection with the helminth Schistosoma mansoni, parasite eggs elicit a Th cell-mediated hepatic granulomatous inflammation that leads to scarring, portal hypertension, hemorrhage, and death. On the other hand, the cytokine IL-10 has been shown to decrease egg Ag-specific Th cell responses by down-regulating MHC class II as well as B7 costimulatory molecule expression on accessory cells. We now test the effect of IL-10 in vivo on models of egg Ag-specific hypersensitivity as well as on the natural disease. Systemic administration of IL-10 significantly inhibited delayed-type hypersensitivity reactions to egg Ag as well as primary and secondary granuloma formation to eggs embolized in the lung. However, significant inhibition of hepatic granuloma formation associated with the natural infection required the use of an IL-10/Fc fusion protein with a prolonged in vivo half-life. Lymph node cells from IL-10/Fc-treated mice produced less IL-2 and IFN-gamma, and more IL-4 and IL-10 than control cells, suggesting that reduced egg granuloma formation resulted primarily from down-regulation of Th1 responses. These results indicate that suitable administration of exogenous IL-10 can be effective in ameliorating immunopathologic damage associated with schistosomiasis.

Animals↗

PubMed1998

Chorionic morphology of eggs of the Psorophora confinnis complex in the United States. II. Pre- and postdeposition studies of Psorophora Columbiae (Dyar and Knab) eggs.

Scanning and transmission electron microscopic techniques were used to determine the morphology of the exochorion and endochorion of Psorophora columbiae eggs as found in the ovary and after deposition. In the ovary, the outer chorionic reticulations and tubercles of the eggs are ornately developed below the follicular epithelium. Expansion of the eggs after deposition causes the outer chorionic reticuli and sculpturing (i.e., minor and major tubercles) to form the characteristic spinose appearance of Psorophora eggs. Transmission electron microscopic micrographs of cross and sagittal sections of the chorionic layers of Ps. columbiae eggs indicate that the major outer chorionic tubercles have 3 distinct regions. The 3rd region was found only on the dorsum of the tubercle. Elevation of major tubercles appears to be regulated, at least in part, by ambient moisture conditions, and one function of these tubercles seems to be adhesion of the egg to an oviposition surface.

Animals↗

PubMed2003

Assessment of egg nutrient compositional changes and residue in eggs, tissues, and excreta following oral administration of atorvastatin to laying hens.

Laying hens were fed a control diet alone or with 0.06 g of atorvastatin, a synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, per 100 g of diet for 20 days. Compared to controls, egg yolks from treated hens contained greater amounts of amino acids and reduced levels of total fatty acids and cholesterol. In contrast, egg albumen amino acid contents were unaffected by dietary treatments. In a residue study, seven hens each received a single oral dose of approximately 20 microCi of [(14)C]atorvastatin. Approximately 71% of the radioactivity was recovered in the excreta and liver, whereas virtually no radioactivity was detected in kidney, heart, muscle, bile, plasma, or egg albumen at 15 days postdosing. Yolk radioactivity peaked at 4 days postdosing in six of the seven birds and was absent in eggs laid after day 10. Reminiscent of that of certain antibiotic drugs, the atorvastatin egg residue pattern appeared to coincide with the physiological pattern of daily yolk accretion within the ovary.

Amino Acids↗

PubMed2002

Distribution of aldrin and its epoxide (dieldrin) in egg-forming tissues and eggs of laying hens following an oral application.

Laying hens were treated orally with a single dose of aldrin (1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5,8-dimethanonaphthalene, AD) 1 mg kg(-1) bw. Concentrations (microg g(-1)) of AD or its epoxide, dieldrin (1,2,3,4,10,10-hexachloro-6,7-epoxy-1,4,4a,5,6,7,8,8a-octahydroendo-,exo-1,4:5,8-dimetha-nonaphthalene, DD), in the main tissues involved in egg formation (blood, liver, ovary, and oviducts) and egg yolk, collected at 1 day after AD dosing, were determined by normal-phase high-performance liquid chromatography. The limits of determination were 0.07 microg g(-1) for AD and 0.08 microg g(-1) for DD, respectively. In extractable fats from the above tissues and egg yolk, AD was found in the egg yolk; however, no AD was found in tissues involved in egg formation. DD was found in all tissues examined here. The DD level was highest in the liver and was lowest in the blood (P<0.01). These results suggest that the epoxidation of AD to DD occurred rapidly in the hen.

Administration, Oral↗

PubMed1987

Allergenic cross-reactivity of egg-white and egg-yolk proteins. An in vitro study.

The radioallergosorbent test (RAST) and RAST inhibition test were used to examine cross-allergenicity amongst the major hen's egg-white and egg-yolk proteins. Using ovalbumin as a reference allergen to compare cross-reactivity, it was apparent that the proteins conalbumin, ovomucoid and lysozyme substantially inhibited binding to ovalbumin discs of IgE in the sera of patients clinically hypersensitive to egg. The converse situation with conalbumin, ovomucoid and lysozyme on the discs and ovalbumin as the inhibitor also resulted in significantly decreased levels of IgE binding to the proteins on the discs. It was also demonstrated that cross-reactions occurred between ovalbumin and the yolk protein, apovitellenin I. Cross-reaction was also observed surprisingly when egg lysozyme was on the disc and the milk protein allergen alpha-lactalbumin was used as the inhibitor. The demonstration of cross-reaction between all of these proteins may signify that there are a number of common allergenic determinants on these egg proteins, thus providing a molecular basis for the phenomenon of cross-reactivity.

Adolescent↗

PubMed1983

Heterogeneity of serum, egg amylase and egg lysozyme in the interspecific hybrids of ducks.

Polymorphism of serum and egg amylase by means of horizontal agarose gel electrophoresis and egg lysozyme by means of horizontal starch gel electrophoresis in Pekin, Muscovy ducks and their interspecific hybrids was studied. In the interspecific hybrids of ducks the codominant type of heredity of serum, egg yolk and egg white amylase isozymes, as well as egg white lysozyme, were found.

Amylases↗