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PubMed2004

Investigating the eggshell conductance and embryonic metabolism of modern and unselected domestic avian genetic strains at two flock ages.

The objective of this study was to determine if broiler strain and breeder flock age affect eggshell conductance, fertility, and hatchability parameters; heart and hepatic glycogen concentrations at hatch; and embryonic metabolism throughout incubation. The 3 broiler strains investigated were HBY, a modern commercial broiler strain selected for high breast yield; WBM, a modern commercial broiler strain selected for the whole bird market; and UN78, a female broiler parent strain unselected since 1978. Fertility and hatchability parameters for each of the 3 strains were determined when the flocks were 32, 34, 37, and 38 wk of age. Eggshell conductance was measured on separate eggs produced from flocks at 37, 45, and 53 wk of age. Concurrently, fertile hatching eggs from the 3 broiler strains at 2 flock ages (33 and 38 wk) were incubated in individual metabolic chambers. Total daily CO2 production of each embryo was measured. Strain and flock age did not influence any of the fertility or hatchability parameters. Strain had no effect on conductance, but eggs from the 37-wk-old flocks had higher conductance than eggs from the 45- or 53-wk-old flocks, which did not differ from one another. Strain had no significant effect on average total CO2 production over the entire 21.5 d of incubation. However, embryos from the 38-wk-old flock produced more total CO2 than did embryos from the 33-wk-old flocks. Also, there was an interaction between strain and flock age for total CO2 production; UN78 embryos from the 33-wk flocks had higher CO2 production than WBM embryos, and the CO2 production of HBY did not differ from either strain. When embryos from the 38-wk flocks were compared, WBM embryos had higher CO2 production than did UN78 embryos, and HBY embryos did not differ from either strain. The data showed that in the 3 strains examined in this study, genetic differences in embryonic metabolic rate were dependent upon breeder flock age.

Age Factors↗

PubMed1992

Pleistocene geochronology and palaeothermometry from protein diagenesis in ostrich eggshells: implications for the evolution of modern humans.

Proteinaceous residues incorporated within the crystal structure of ostrich eggshells (OES) are retained without loss over geological time exceeding 10 million years. Degradation of the polypeptides, including hydrolysis to smaller peptide fragments and eventual release of free amino acids, decomposition, and racemization and epimerization occur at regular, predictable rates dependent on ambient temperature. The extent of isoleucine epimerization (aIle/Ile ratio) in OES follows linear first-order reversible kinetics in controlled-temperature laboratory simulations of time up to an aIle/Ile ratio in excess of 1.0. The hydrolysis of leucine also follows a predictable pattern, but deviates from first-order kinetics. A nonlinear mathematical model has been developed that adequately describes the pattern of leucine hydrolysis through a wide temperature range. Arrhenius parameters were derived from laboratory experiments combined with rate constant values found for 14C-dated OES from stratified caves in southern Africa. These parameters for isoleucine epimerization and leucine hydrolysis differ by ca. 10%, allowing the simultaneous solution of the two equations for temperature, independent of sample age. Although the uncertainty of the simultaneous temperature is relatively high (+/- 10 degrees C), it provides an effective means of identifying burned samples. If sample age is known, palaeotemperatures (the integrated thermal history experienced by an eggshell as opposed to an 'instantaneous' temperature) can be calculated with a precision of better than +/- 1 degrees C. The ages of levels at Border Cave, South Africa, from which anatomically modern human skeletal remains have been recovered, are dated by the extent of isoleucine epimerization in associated OES.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Cloning and analysis of the dec-1 female-sterile locus, a gene required for proper assembly of the Drosophila eggshell.

Female-sterile mutations at the dec-1 (defective chorion-1) locus of Drosophila severely disrupt the organization of the eggshell late in oogenesis. Previous characterization of dec-1 mutations has correlated the defects with failure to accumulate an early eggshell protein that undergoes proteolytic cleavage during choriogenesis. To enable further study of the regulation and processing of dec-1 products, we have molecularly cloned the locus and characterized its transcripts. Chromosome jumping was used to isolate a deficiency breakpoint within the locus. Overlapping genomic clones from a wild-type library were then obtained, and a region including the dec-1 locus was identified by hybridization to cDNA probes complementary to RNA from stage 9-10 egg chambers. Analysis of genomic rearrangements associated with the locus verified its identity. Two transcripts from the locus have been identified and characterized using cDNA clones, RNase protection, and primer extension analyses. A 4.0-kb transcript accumulates maximally in stages 9-10, when the primary follicle cell protein associated with dec-1 mutations is synthesized. A second transcript of 5.8 kb, generated by alternative splicing, accumulates during stages 11-12. These results are discussed in light of previous analysis of dec-1 mutations.

Animals↗

PubMed1978

Isolation of messenger RNA coding for eggshell protein of the DNA-eliminating nematode Ascaris lumbricoides.

Poly(A)-containing RNA from polyploid uterine epithelial cells of Ascaris lumbricoides has been isolated by poly(U)-Sepharose chromatography. The bulk of poly(A)-containing RNA migrates as 18-S RNA in formamide/polyacrylamide gels. In a cell-free wheat germ system, this RNA directs the synthesis of a polypeptide with identical migration behavior in dodecylsulphate/urea/polyacrylamide gels as the polypeptide isolated from the proteinaceous eggshell. The two proteins reveal almost identical peptide patterns in fingerprint analysis. The authentic eggshell protein has been identified as a glycoprotein with a molecular weight of about 10000, as determined by dodecylsulphate/polyacrylamide gel electrophoresis. The apparent discrepancy between mRNA length and the required coding length for the protein is discussed.

Animals↗

PubMed1983

The inhibitory effect of some chlorinated hydrocarbon pesticides on the ATP-dependent Ca2+ binding of the particulate fraction of the eggshell gland mucosa cells.

The pesticide p-p'-DDT and its persistent metabolite p-p'-DDE cause thinning of the eggshells in several species of birds. In earlier investigations on ducks this thinning was found to be associated with a reduction of the ATP-dependent Ca2+ binding to a homogenate of the shell gland mucosal cells by DDE. The activity of a Ca2+-Mg2+-activated ATPase in the homogenate was also decreased on administration of DDE in vivo. We have therefore investigated the in vitro effects of some other chlorinated hydrocarbon pesticides of ecotoxicological interest on the ATP-dependent Ca2+ binding and the Ca2+-Mg2+-activated ATPase activity in a homogenate of the eggshell gland mucosa of the hen and determined the molar concentrations that produced 50% inhibition (=IC50). Several of the investigated compounds, namely toxaphene, chlordane, p-p'-DDD, o-p'-DDE, p-p'-DDT, methoxychlor and PCB (Arochlor 1242), had a similar IC50 to inhibit the Ca2+ binding as p-p'-DDE. Lindane, p-p'-DDA and biphenyl had an IC50 3.3-4 times higher and that of 2.4 D was 13.5 times higher than that of p-p'-DDE. When the IC50 of some of the compounds (p-p'-DDE, PCB, toxaphene, Lindane) was determined that decreased the Ca2+-Mg2+-activated ATPase of the homogenate it was found to be only 18 to 29 per cent of that needed to inhibit the Ca2+ binding by the homogenate. It is therefore probable that some other effect than inhibition of this enzyme is also involved in the Ca2+-binding process and affected by the compounds.

Adenosine Triphosphatases↗

PubMed1986

Effect of some metal compounds on the Ca2+ binding and Ca2+-Mg2+-ATPase activity of eggshell gland mucosa homogenate from the domestic fowl.

Some metal ions, e.g. Hg2+, Cd2+ and Al3+, can have the effects as ecotoxicological agents, of causing eggshell thinning and breakage in birds. In a homogenate of the Ca2+-secreting part of the eggshell gland mucosa, a study was made of the influence of Hg2+, Cd2+, Cu2+, Pb2+, methyl-Hg+, Zn2+, V3+, Al3+ and Ni2+ in different concentrations on the rate of ATP-dependent 10(-4) M Ca2+ binding. All compounds had an inhibitory action. The most potent metal (Hg2+) produced 50% inhibition (IC50) at 1.1 X 10(-6) M, whereas this value for the least potent compound (Ni2+) was 9 X 10(-4) M. The specific Ca2+-Mg2+-ATPase activity was also inhibited by the tested metal ions. In all cases except methyl-Hg+ the IC50 for this activity was lower than that for Ca2+ binding. The most potent ion in this respect was Cd2+, with an IC50 of 8 X 10(-8) M, and the least potent was methyl-Hg+, with an IC50 of 1.4 X 10(-3) M. Pb2+ and Cd2+ in a concentration range of 10(-5)-10(-4) stimulated the Mg2+-ATPase activity, however, to almost the same extent as 10(-4) M Ca2+. A possible explanation for this effect is that these ions may have an affinity for sites of Ca2+ binding of the polypeptide calmodulin and thereby influence the Ca2+ metabolism of the shell gland mucosa.

Animals↗

PubMed1971

DDE residues and eggshell changes in Alaskan falcons and hawks.

Eggshell thickness after exposure to DDT was reduced by 21.7 percent in Alaskan tundra peregrines, by 16.8 percent in taiga peregrines, by 7.5 percent in Aleutian peregrines, by 3.3 percent in rough-legged hawks, and not at all in gyrfalcons. Tundra peregrine eggs contain an average of 889 parts of DDE per million (lipid basis); taiga peregrine eggs contain 673 parts per million; Aleutian peregrine eggs contain 167 parts per million; rough-legged hawk eggs contain 22.5 parts per million; and gyrfalcon eggs contain 3.88 parts per million. These changes in eggshell thickness and the pesticide residues reflect different degrees of exposure to contamination. There is a highly significant negative correlation between shell thickness and DDE content in peregrine eggs. Tundra and taiga peregrines have fledged progressively fewer young each year since 1966.

Alaska↗

PubMed2002

Mechanical strain regulation of the chicken glypican-4 gene expression in the avian eggshell gland.

Comparison of RNA fingerprinting of the avian eggshell gland (ESG) without and with an egg revealed upregulation of a 382-bp cDNA fragment that showed high homology to the mammalian glypican 4 (GPC-4). The gene sequence revealed a conserved glypican signature, a glycosyl phosphatidyl inositol-anchorage site, and cystein residues, most of which were conserved. GPC-4 was expressed in the ESG in a circadian fashion only during the period of eggshell calcification, when maximal mechanical strain was imposed. Removal of the egg just before to its entry into the ESG, with consequent elimination of the mechanical strain, caused reduction in the gene expression. Artificial application of the mechanical strain induced expression of the GPC-4 gene that was related to the level of the strain. GPC-4 expression was strain dependent in other parts of the oviduct. In the ESG, GPC-4 was expressed exclusively by the glandular epithelium and not by the pseudostratified epithelium facing the lumen. In summary, we cloned the avian homologue of GPC-4, established its pattern of expression in the avian ESG, and demonstrated for the first time that this gene is regulated by mechanical strain.

Amino Acid Sequence↗

PubMed1982

Changes in eggshell conductance after transfer of hens from an altitude of 3,800 to 1,200 m.

Hens acclimated to an altitude of 3,800 m (PB 480 Torr) were transferred to 1,200 m (PB 657 Torr). Eggs were collected before departure and daily after the transfer so that changes in eggshell conductance could be studied. Over the next 2 mo eggshell conductance increased 30%, presumably to compensate for the 37% reduction (from 657 to 480 Torr) in gas diffusivity at the lower altitude. Measurements of shell thickness and number of pores in the shell allow one to calculate that most of the change in total pore area occurred by an increase in cross-sectional area of individual pores.

Adaptation, Physiological↗

PubMed2003

Eggshell and egg yolk proteins in fish: hepatic proteins for the next generation: oogenetic, population, and evolutionary implications of endocrine disruption.

The oocyte is the starting point for a new generation. Most of the machinery for DNA and protein synthesis needed for the developing embryo is made autonomously by the fertilized oocyte. However, in fish and in many other oviparous vertebrates, the major constituents of the egg, i.e. yolk and eggshell proteins, are synthesized in the liver and transported to the oocyte for uptake. Vitellogenesis, the process of yolk protein (vitellogenin) synthesis, transport, and uptake into the oocyte, and zonagenesis, the synthesis of eggshell zona radiata proteins, their transport and deposition by the maturing oocyte, are important aspects of oogenesis. The many molecular events involved in these processes require tight, coordinated regulation that is under strict endocrine control, with the female sex steroid hormone estradiol-17beta in a central role. The ability of many synthetic chemical compounds to mimic this estrogen can lead to unscheduled hepatic synthesis of vitellogenin and zona radiata proteins, with potentially detrimental effects to the adult, the egg, the developing embryo and, hence, to the recruitment to the fish population. This has led to the development of specific and sensitive assays for these proteins in fish, and the application of vitellogenin and zona radiata proteins as informative biomarkers for endocrine disrupting effects of chemicals and effluents using fish as test organisms. The genes encoding these important reproductive proteins are conserved in the animal kingdom and are products of several hundred million years of evolution.

Journal Article↗

PubMed1997

Two signalling pathways specify localised expression of the Broad-Complex in Drosophila eggshell patterning and morphogenesis.

The Drosophila eggshell, which has a pair of chorionic appendages (dorsal appendages) located asymmetrically along both the anterior/posterior and dorsal/ventral axes, provides a good model to study signal instructed morphogenesis. We show that the Broad-Complex, a gene encoding zinc-finger transcription factors, is essential for the morphogenesis of dorsal appendages and is expressed in a bilaterally symmetrical pattern in the lateral-dorsal-anterior follicle cells during late oogenesis. This is induced and specified along the dorsoventral axis by an epidermal growth factor receptor signalling pathway, which includes a localised transforming growth factor-alpha like molecule, Gurken, in the oocyte and the Drosophila EGF receptor homologue, Torpedo, in the surrounding somatic follicle cells. Furthermore, the precisely localised expression of BR-C along the AP axis requires a separate signalling pathway, initiated by a transforming growth factor-beta homologue, Decapentaplegic, in nearby follicle cells. These two signalling pathways, one from the oocyte and the other from the follicle cells, co-ordinately specify patches of follicle cells to express the Broad-Complex in a unique position in respect to both major axes, which in turn directs the differentiation of the dorsal appendages in the correct position on the eggshell.

Animals↗

PubMed2001

Specific domains drive VM32E protein distribution and integration in Drosophila eggshell layers.

A study was made of the localization and assembly of the VM32E protein, a putative vitelline membrane component of the Drosophila eggshell. The results highlight some unique features of this protein compared with the other proteins of the same gene family. At the time of its synthesis (stage 10), the VM32E protein is not detectable in polar follicle cells. However, it is able to move in the extracellular space around the oocyte and, by stage 11 is uniformly distributed in the vitelline membrane. During the terminal stages of oogenesis the VM32E protein is partially released from the vitelline membrane and becomes localized in the endochorion layer also. By analyzing transgenic flies carrying variously truncated VM32E proteins, we could identify the protein domains required for the proper assembly of the VM32E protein in the eggshell. The highly conserved vitelline membrane domain is implicated in the early interactions with other components and is required for cross-linking VM32E protein in the vitelline membrane. The terminal carboxylic domain is necessary for localization to the endochorion layer. Protein with the C-end domain deleted is localized solely to the vitelline membrane and cross-linked only in laid eggs, as occurs for the other vitelline membrane proteins.

Amino Acid Sequence↗

PubMed2005

Oxygen and water flux across eggshells of Manduca sexta.

Insect eggs must obtain oxygen across the eggshell to support embryonic development. Because eggs are small, obtaining enough oxygen would seem trivial. Recent work, however, has shown that eggs of a moth, Manduca sexta, are oxygen limited at high but realistic temperatures (32-37 degrees C) and that P(O2) drops steeply across the eggshell. Here we use theoretical and experimental approaches to partition the total resistance to oxygen flux among several steps in the oxygen cascade from environment to embryo. Standard mass-transfer analysis suggests that boundary layers of air around eggs, and around substrates to which they are attached, offer negligible resistance. Likewise, a mathematical model, parameterized using published and newly obtained morphological data, predicts that air-filled parts of the chorion also do not resist oxygen flux. This prediction was confirmed by experiments that measured rates of carbon dioxide emission from batches of eggs subjected simultaneously to hypoxia and inert gas substitution: depression of metabolic rate by hypoxia was not rescued when the diffusion coefficient of oxygen in air was doubled by substituting helium for nitrogen. The model did predict, however, that a set of subchoral layers (a crystalline chorionic layer, a wax layer and the vitelline membrane) could account for most or all of the total resistance to oxygen flux. Support for this prediction was obtained from two sequential experiments. First, eggs extracted with chloroform:methanol had highly elevated rates of water loss, suggesting that indeed eggs of M. sexta are waterproofed by wax. Second, rates of water loss and carbon dioxide emission from batches of eggs, measured from laying to hatching, changed in parallel over development. These data suggest that a single layer, likely a wax layer or a combination of wax and other subchoral layers, provides the main resistance to water efflux and oxygen influx.

Animals↗

PubMed2002

Crystallization and preliminary x-ray analysis of ovocleidin-17 a major protein of the gallus gallus eggshell calcified layer.

In this work, we report the crystallization of ovocleidin-17, the major protein of the avian eggshell calcified layer and the preliminary X-ray characterization of this soluble protein which is implied into the CaCO(3) formation of the eggshell in avians. Crystals belong to one of the trigonal space group P3 with cell dimensions a= b= 59.53 A and c = 83.33 A, and alpha=beta= 90 degrees and gamma=120 degrees. Crystals diffract up to 3.0 A.

Animals↗

PubMed1993

Effect of concentration of mixed-function oxidase on concentration of estrogen, rate of egg lay, eggshell thickness, and plasma calcium in laying hens.

The objective of this study was to investigate the effect of induction or depression of mixed-function oxidases (MFO) by xenobiotics on the peripheral concentration of estrogens, plasma Ca, rate of egg lay, and eggshell thickness in laying hens. In Exp. 1, 100 mg of phenobarbital (PB) administered orally each day for 3 or 7 d reduced concentrations of estrone and estradiol (E2) in serum. In Exp. 2, 25, 50, or 100 mg of PB was administered orally to laying hens for 3 or 7 d. Treatment with 100 mg of PB for 3 d or with 25, 50, or 100 mg for 7 d significantly increased liver:body weight ratios. Microsomal protein increased after 7 d of 50 or 100 mg of PB administration. Feeding PB decreased concentration of E2 and total plasma Ca in a dose- and period-dependent manner. Concentration of E2 was reduced to 10% of control, whereas hepatic cytochrome P-450 increased significantly with dose. The correlation between concentration of E2 and P-450 was negative and significant. Total Ca in the plasma was highly, positively correlated with concentration of E2. Eggshells were thinner from hens treated with 100 mg of PB for both 3 or 7 d than those from control hens. Rate of egg lay was reduced by 100 mg of PB for 7 d. In Exp. 3, .5 mL of CCl4 given orally for 1 d decreased P-450 and increased E2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Ovalbumin is a component of the chicken eggshell matrix.

The protein components of biomineralized structures (matrix proteins) are believed to modulate crystal nucleation and growth, and thereby influence the shape and strength of the final structure. The chicken eggshell contains a complex array of distinct matrix proteins. One of these was found to have similar molecular weight and chromatographic properties as purified egg ovalbumin. A commercially available antibody to ovalbumin was utilized for western blotting to demonstrate that ovalbumin is one of the matrix proteins this is extracted from decalcified eggshell. Immunohistochemistry revealed that ovalbumin is found only in the mammillary bodies of decalcified shell, and is not distributed throughout the shell matrix. These results indicate that ovalbumin is present during the initial phase of shell formation and becomes incorporated into the protein matrix of the mammillary bodies. However, it is not yet clear whether ovalbumin at this site plays a specific role in shell mineralization.

Animals↗

PubMed1993

Influence of large doses of ascorbic acid on performance, plasma calcium, bone characteristics, and eggshell quality in broilers and Leghorn hens.

Four experiments were conducted using broilers (Experiments 1 and 2) or White Leghorn hens (Experiments 3 and 4) to determine the effects of large doses of dietary ascorbic acid on performance, plasma concentration of total and ionic calcium, bone characteristics, and eggshell quality. A total of 564 male broilers were fed diets containing ascorbic acid (ranging from 0 to 3,000 ppm) from 3 to 7 wk of age. Weight gain and feed conversion were measured, and blood plasma was analyzed for total and ionic calcium. Leg bones (femur, tibia, and metatarsus) were analyzed for bone mineral content, density, and breaking strength. In Experiments 3 and 4, a total of 484 Leghorn hens were fed diets containing ascorbic acid (ranging from 0 to 3,000 ppm) for 4 wk. Egg weight and specific gravity were determined, and plasma and tibiae were analyzed as in Experiments 1 and 2. Results of the broiler experiment (Experiment 2) indicated that plasma ionic calcium was significantly increased (P < .05) in ascorbic-acid-treated birds. Among leg bones examined, femur strength was improved by 16% in birds fed 2,000 ppm of ascorbic acid (Experiment 2). Other bone characteristics were not affected. In the layer experiments (Experiments 3 and 4), egg weight increased up to 5% and egg specific gravity was improved in hens fed 2,000 or 3,000 ppm of ascorbic acid, which also had increased calcium in the blood. Results suggest that large doses of ascorbic acid in the diet influence calcium metabolism, affecting bone and eggshell mineralization in chickens.

Animals↗

PubMed1994

Effects of dietary thiouracil on thyroid activity, egg production, and eggshell quality in commercial layers.

The effects of thyroid suppression induced during the rearing period by providing various dietary thiouracil (TU) regimens on plasma thyroxine (T4) concentrations, growth, and subsequent egg production (EP) and eggshell quality were determined in Single Comb White Leghorn chickens. Thiouracil was provided in the feed at levels of 0, .1, and .2% from 0 to 6 wk of age in Experiment 1, and at levels of 0, .05, and .1% from 6 to 16 wk of age in Experiment 2. In both experiments, T4 concentrations were reduced during TU treatment. However, T4 later became elevated at 12 and 20 wk in both dosage level groups in Experiment 1. Additionally, BW and egg weights were suppressed by both TU treatments, and EP was reduced up to Week 23 in the .1% TU-treated birds and through Week 25 in the .2% TU-treated birds. No effects on EP were noted in Experiment 2, but feed consumption (FC) was reduced during Week 6 in birds fed .05% TU and during Weeks 6, 10, and 19 in birds fed .1% TU. Both liver and thyroid weights were increased in .1% TU-treated birds relative to controls at Week 16. Eggshell quality was affected only in Experiment 2, in which birds given .05% TU had a higher relative conductance, or maximum rate of water loss, at Week 38 than 0 and .1% TU dosage levels, and .1% TU-treated birds had a higher breaking strength than 0 and .05% TU-treated birds at Week 22.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗