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PubMed2000

Comparative ultrastructure of eggs in Echinostoma paraensei, E. caproni, and E. trivolvis (Trematoda: Echinostomatidae).

The 37-collar-spined echinostomes Echinostoma paraensei, E. caproni, and E. trivolvis are digeneans that live in the intestine of small mammals and birds. Comparative studies of the eggs of these species were done using light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The egg of E. caproni was the largest of the three species studied, whereas the egg of E. trivolvis was the smallest in both length and width. The SEM study showed differences in the aboperculum region of the eggs in all three species. The TEM study showed that the eggshell of all three species consisted of three layers, but no difference in eggshell structure was observed in any species.

Animals↗
PubMed2001

Nematode chitin synthases: gene structure, expression and function in Caenorhabditis elegans and the plant parasitic nematode Meloidogyne artiellia.

Although the presence of chitin in nematodes is well documented little is known about its synthesis in this phyletic group. The recently completed genome sequence of Caenorhabditis elegans predicts two sequences with homology to chitin synthases (chitin-UDP acetyl-glucosaminyl transferase; EC 2.4.1.16). We show that these genes are differentially expressed in a pattern that may reflect different functional roles. One gene is expressed predominantly in the adult hermaphrodite (the main egg-producing stage in the nematode) and later larval stages, which is consistent with a role in production of chitin for the eggshell. The other gene, however, is expressed in the cells that form the pharynx, and only in the period directly preceding a moult. These data suggest that the product of this gene is involved in synthesis of the feeding apparatus, which is replaced during each moult. We have also isolated a full-length genomic sequence of a chitin synthase orthologue from the plant parasitic nematode Meloidogyne artiellia. The single gene present in M. artiellia shows an expression pattern that is consistent with a role for the protein in production of the eggshell.

Animals↗
PubMed1994

The role of vitamin D in chorioallantoic membrane calcium transport.

1,25-Dihydroxyvitamin D3 (1,25-(OH)2D3) is essential for the transport of eggshell calcium to the embryo across the chorioallantoic membrane (CAM). CAM contains the vitamin D receptor that increases following 1,25-(OH)2D3 injection into embryos at day 10 of incubation. Further, a single injection of 100 ng of 1,25-(OH)2D3 into vitamin D-deficient quail eggs at day 10 of incubation resulted in a significant increase in both body and yolk calcium. This is accompanied by an increase in carbonic anhydrase from low levels in deficiency to normal levels. Acetazolamide (AZ), a specific carbonic anhydrase inhibitor injected into the quail embryos, caused hypocalcemia and hyperphosphatemia. This is similar to the hypocalcemia and hyperphosphatemia found in vitamin D-deficient embryos. These results suggest that one mechanism of action of vitamin D in the mobilization of eggshell calcium is the activation of carbonic anhydrase that acidifies the calcium carbonate shell.

Acetazolamide↗
PubMed1983

Isolation of germ line-dependent female-sterile mutation that affects yolk specific sequestration and chorion formation in Drosophila.

Two loci on the X chromosome have been implicated in choriogenesis by in situ hybridization of poly A-containing RNA from choriogenic eggchambers to Drosophila polytene chromosomes (A.C. Spradling and A.P. Mahowald (1979). Cell 16, 589-598): 7E and 12E. At least two genes coding for major eggshell proteins map to region 7E (A.C. Spradling, M.E. Digan, A.P. Mahowald, M. Scott, and E.A. Craig (1980). Cell 19, 905-914). In an effort to elucidate the functional role of the 12E gene product, 3600 EMS-treated X chromosomes were screened for recessive female-sterile mutations that mapped within the region 11F10-12F1. Four independent female-sterile mutations were recovered, three of which fell into one complementation group (fs29, fs117, and fs445). Mapping by analysis of recombinant progeny as well as of trans heterozygotes utilizing other deficiency chromosomes showed that the three noncomplementing mutations all mapped to region 12E1-12F1. Studies comparing chorion morphology and protein synthesis indicate localized perturbations in the extracellular assembly of eggshell components in mutant eggchambers. The germ line dependence of the mutations was established using germ line mosaics constructed by pole cell transplantation. Analysis of eggchamber protein accumulation patterns showed reduced amounts of yolk polypeptides (YPs) in the mutants. The elevated concentrations of YPs found in mutant hemolymph coupled with the normal YP biosynthetic patterns and active uptake of trypan blue by mutant oocytes suggest that 12E sequences play a role in yolk-specific sequestration.

Animals↗
PubMed1984

Localization of a gene for a minor chorion protein in Drosophila melanogaster: a new chorion structural locus.

A minor chorion protein (called s70) with an approximate molecular weight of 70,000 D has been characterized in Drosophila melanogaster. The Staket geographic strain was found to carry an electrophoretic variant of this eggshell component and was used to determine the chromosomal location of the s70 gene. Our results establish a new locus for a chorion gene near yellow on the X chromosome and represent the first mapping of a quantitatively minor eggshell protein.

Animals↗
PubMed1991

Vitamin D and chick embryonic yolk calcium mobilization: identification and regulation of expression of vitamin D-dependent Ca2(+)-binding protein, calbindin-D28K, in the yolk sac.

The developing chick embryo acquires calcium from two sources. Until about Day 10 of incubation, the yolk is the only source; thereafter, calcium is also mobilized from the eggshell. We have previously shown that during normal chick embryonic development, vitamin D is involved in regulating yolk calcium mobilization, whereas vitamin K is required for eggshell calcium translocation by the chorioallantoic membrane. We have studied here the biochemical action of 1,25-dihydroxy vitamin D3 in the yolk sac by examining the expression and regulation of the cytosolic vitamin D-dependent calcium-binding protein, calbindin-D28K. Two types of embryos are used for this study, normal embryos developing in ovo and embryos maintained in long-term shell-less culture ex ovo, the latter being dependent solely on the yolk as their calcium source. Our findings are (1) calbindin-D28K is expressed in the embryonic yolk sac, detectable at incubation Days 9 and 14; (2) the embryonic yolk sac calbindin-D28K resembles that of the adult duodenum in both molecular weight (Mr 28,000) and isoelectric point, as well as the presence of E-F hand Ca2(+)-binding structural domains; (3) systemic calcium deficiency caused by shell-less culture of chick embryos results in enhanced expression of calbindin-D28K in the yolk sac during late development; (4) yolk sac calbindin-D28K expression is inducible by 1,25-dihydroxy vitamin D3 treatment in vivo and in vitro; and (5) immunohistochemistry revealed that yolk sac calbindin-D28K is localized exclusively to the cytoplasm of the yolk sac endoderm. These findings indicate that the chick embryonic yolk sac is a genuine target tissue of 1,25-dihydroxy vitamin D3.

Animals↗
PubMed1986

Effects of suppression and resumption of shell formation and parathyroid hormone on uterine calcium-binding protein, carbonic anhydrase activity, and intestinal calcium absorption in hens.

The calcium absorption and duodenal and uterine vitamin D-dependent calcium-binding protein (CaBP-28K) levels were decreased in hens when eggshell calcification was suppressed by premature expulsion of the egg. Nevertheless, these levels remained higher than those of immature pullets or pullets treated with estrogen. The resumption of shell formation by hens which had previously laid soft-shell eggs was associated during calcification of the first egg with increases in intestinal Ca absorption. CaBP concentration, and alkaline phosphatase activity. The increase in uterine CaBP concentration preceded the stage of rapid calcium deposition. Uterine carbonic anhydrase activity was increased by sexual maturity but not consistently by shell formation. Ablation of the parathyroids just before the resumption of shell formation suppressed the increases in duodenal calcium absorption and CaBP concentration elicited by egg calcification. In contrast, the increase in CaBP level was maintained in the uterus of parathyroidectomized hens, in spite of the decreased shell deposition. Previous studies indicated that increased uterine CaBP associated with eggshell calcification is not elicited by vitamin D. The present study confirms this observation and also shows that these changes are not elicited by either PTH or sex steroid hormones.

Animals↗
PubMed1979

Water-vapor conductance of testudinian and crocodilian eggs (class reptilia).

Flexible-shelled eggs of snapping turtles (Chelydra serpentina) have conductances to water vapor that are 55 times higher than predicted for avian eggs of similar size, whereas rigid-shelled eggs of softshell turtles (Trionyx spiniferus) and American alligators (Alligator mississippiensis) have conductances that are only five times higher than expected for comparable eggs of birds. The differences between empirical and predicted values result from the much higher effective pore areas in reptilian eggshells than in those of birds. The relatively high porosities of these reptilian eggs presumably facilitate the transport of oxygen and carbon dioxide eggshells in later stages of incubation when air trapped inside nest chambers may become hypoxic and hypercapnic, yet seem not to lead to excessive transpiration of water vapor owing to the high humidities in nests where incubation occurs.

Alligators and Crocodiles↗
PubMed1982

The initiation of pulmonary respiration in a bird embryo: blood and air cell gas tensions.

The gaseous stimuli for pipping and the initiation of pulmonary respiration were examined in th Wedge-tailed Shearwater (Puffinus pacificus) by measuring blood gas tensions in the chorioallantoic vein and air cell gas tensions (Pao2 and Paco2) prior to and during the pipping process. External pipping (shell fracture) was the first observed pipping event and preceded internal pipping (penetration of air cell and initiation of pulmonary ventilation). This sequence of cracking the eggshell prior to the initiation of pulmonary respiration resulted in higher Pao2 and lower Paco2 values than the pre-pip air cell gas tensions. Pip-cracks in the shell allow greater diffusion than could normally be obtained across the intact eggshell. This also favors the establishment of large partial pressure gradients of O2 and CO2 across the chorioallantois, thereby improving gas exchange across the inner resistance barrier. The gaseous stimuli for pipping may be attenuated and thus favor the 5-6 day pip-to-hatch interval in this species.

Allantois↗
PubMed1988

Development of the chick embryo: effects of egg mass.

We asked to what extent, within a species, differences in egg mass, hence in eggshell surface area and O2 conductance, could affect the development of the avian embryo. Large (L, about 70 g) and small (S, about 55 g) fertile chicken eggs were simultaneously incubated and the embryos studied at day 18, i.e. before the onset of lung ventilation. Embryo mass and O2 consumption (VO2) (measured with a manometric technique) normalized per mass of the freshly laid egg were higher in S than in L, while no differences occurred after normalization by egg surface area (S.A.). Egg water vapour conductance, which is proportional to O2 conductance, was also found to be directly proportional to egg S.A. Hence, the mass and VO2 of the embryo are more closely related to the O2 conductance of the eggshell than to egg mass, giving support to the concept that the avian embryo's VO2 is not an invariable species characteristic, but a variable dependent on O2 availability. Hatching and viability did not differ between L and S, and the specific mass of heart and lung and their cellular (DNA) concentration were also similar between the two groups of embryos. Therefore, differentiation of tissues and organogenesis do not seem to be affected by the differences in total O2 availability determined by the differences in egg mass, while they are probably more important than total tissue mass in setting the time of hatching.

Animals↗
PubMed1994

A new member to the astacin family of metalloendopeptidases: a novel 1,25-dihydroxyvitamin D-3-stimulated mRNA from chorioallantoic membrane of quail.

1,25-Dihydroxyvitamin D-3 is essential for the utilization of eggshell calcium by avian embryo through the chorioallantoic membrane (CAM). A cDNA library was constructed from poly(A)+ RNA extracted from vitamin D-deficient CAMs given 1,25-dihydroxyvitamin D-3. Screening this library by differential hybridization yielded a full-length (approximately 1.8 kb) cDNA, whose corresponding mRNA is increased 3-fold 2.5 h after a single injection of 1,25-(OH)2D3. The complete nucleotide sequence for the full-length cDNA has been determined. An open-reading frame, corresponding to a 310 amino acid, 41 kDa protein was found. Searching protein sequence data bases revealed a strong similarity to the following proteases: astacin, a crayfish digestive protease, Oryzias latipes hatching enzyme constituent protease (Orz), Xenopus laevis developmentally regulated UVS.2 protein secreted by the hatching gland of embryos, the NH2-terminal domain of human bone morphogenetic protein (BMP-1) and Drosophila dorsal-ventral patterning tolloid. The cDNA has approximately 36% overall identity with astacin and BMP-1, and is more than 60% identical to either Orz or UVS.2. Moreover, multiple alignment analysis indicates that 37 residues, including 3 cysteine residues, are strictly conserved in the complete 200-amino acid astacin sequence. All 6 proteins contain a zinc-binding motif (HEXXH), found at the active site of most metalloendopeptidases. This motif is found within an extended sequence of HEXXHXXGFXHE that is unique to this subgroup of metalloendopeptidases. In addition, the 6 proteins have 50% identity (including the present cDNA) and 79% are conserved in 4 of these proteins in a 24-amino acid sequence that includes the putative active site. The level of mRNA for the new protein reaches a maximum at day 12 of embryonic life and declines thereafter. It is suggested that this clone corresponds to an mRNA encoding for a protease that may play a role in the degradation of eggshell matrix.

Allantois↗
PubMed1984

Changes in activity of chicken medullary bone cell populations in relation to the egg-laying cycle.

Osteoblastic and osteoclastic activity was studied in avian medullary bone in vivo. During the active period of eggshell calcification, medullary bone active resorption surface increased ninefold. This correlated with a sevenfold increase in the percentage of active osteoclasts. Osteoblast activity is also increased during the active period, as demonstrated by a twofold increase in the active osteoblastic surface. These findings and our observation that the medullary bone volume remains the same (+/- 13%) whether the eggshell is being formed (active period) or not (inactive period) led to the conclusion that the activities of osteoblasts and osteoclasts rapidly return to balance.

Animals↗
PubMed1989

Blood plasma catecholamine concentration of poult embryos during the transition from diffusive to convective respiration.

1. Plasma catecholamines were determined in turkey embryos. 2. Decreased eggshell permeability depressed dopamine and epinephrine, but increased permeability also depressed epinephrine. 3. Norepinephrine (NE) was greatest before transition to convective respiration when hypoxia occurs. 4. NE was associated inversely with eggshell permeability. 5. Catecholamines appear to assist the embryo during hatching hypoxia.

Animals↗
PubMed1995

Electrophoretical and histochemical characterization of Schistosoma mansoni phenol oxidases.

The phenol oxidase system, which is thought to play a central role in eggshell formation, was analyzed by means of electrophoretic and histochemical techniques. In contrast to current beliefs, our data show that males also express the phenol oxidase activity. The analysis of the electrophoretic pattern from males and females shows differences: adult males present a major band and a faint band, migrating slightly below. Adult females express a more complex pattern composed of four bands. Histochemical studies revealed that female phenol oxidase is concentrated in the vitelline cells, is inhibited by DDC, and this inhibition is correlated with disruption of female egg production. The present studies, in agreement with other reports, reveal that female phenol oxidase may be involved in sclerotization of the S. mansoni eggshell.

Animals↗
PubMed1997

Histological analysis of the egg capsule of the ovoviviparous polystomatid monogenean, Pseudodiplorchis americanus.

Transmission of Pseudodiplorchis americanus is restricted to the brief period when its host, a desert toad, enters water to spawn. The parasite accumulates its entire annual reproductive output within an elongated uterus during the 10-11-month period of host hibernation. Embryos of P. americanus, at all stages of development, are retained within the uterus which eventually becomes packed with around 150 encapsulated infective larvae. Recently formed eggs, which comprise a fertilized ovum and 2-3 vitelline cells, are closely surrounded by a primary eggshell which stains positively for acidic proteins and keratin. Initially, during passage along the proximal uterus, the egg capsule is only 60 microns in diameter, but as it passes to the distal uterus it expands to 800 microns in diameter to accommodate the growing larva. Due to chemical alterations or complete replacement of the shell, the final (secondary) egg capsule is a large sac-like structure composed of elastin. The flexible nature of this shell maximizes the numbers of infective larvae which can be stored in utero. TEM studies have revealed this capsule to be composed of multi-laminate membranes with a specialized cytoplasmic lining involved in a unique mechanism for embryo nutrition. This is the first report of an elastin-type eggshell within the Monogenea.

Animals↗
PubMed2003

Structural and biochemical analysis of the Leptinotarsa decemlineata (Coleoptera; Chrysomeloidea) crystalline chorionic layer.

The developmental aspects of the Leptinotarsa decemlineata crystalline chorionic layer (CCL) morphogenesis, its composition and its supramolecular structure were studied. The mature Leptinotarsa decemlineata eggshell consists of the vitelline membrane and the CCL, while the follicle cell remnants following their degeneration after oogenesis completion constitute the outer chorionic layer. The vitelline membrane and the CCL layers are formed through continuous material deposition from the follicular epithelium, whereas the main morphogenic factor during most insect eggshell formation, namely the follicle cell and oocyte microvilli, are seemingly involved only in vitelline membrane formation. Analysis of the CCL morphogenesis showed that this layer is assembled from a fiber-like pre-crystalline material, which accumulates at the vitelline membrane-follicle cell interface. The mature CCL is about 1 microm thick and exhibits a periodicity of approximately 10 nm, while computer image analysis studies of thin-sectioned CCL revealed the existence of crystalline layers parallel to the CCL surface. Finally, SDS-PAGE-electrophoresis of purified CCLs showed that this crystalline layer is of a proteinaceous nature and is most likely composed of 3-5 polypeptides with a molecular weight ranging in between 28-60 kDa. Overall, these data exemplify for the first time the nature and supramolecular arrangement of a crystalline layer and its constituent molecules in Coleoptera.

Animals↗
PubMed2002

Effect of regional changes to shell conductance on oxygen consumption and growth of chicken embryos.

Diffusive and perfusive gas conductances of the shell and chorioallantois of avian eggs are not homogeneous, leading to disequilibrium between the air cell gas and chorioallantoic venous blood. This study measured the embryonic ability to maintain oxygen consumption rate and growth under experimentally enhanced inhomogeneity of diffusive conductance, achieved by waxing half of the eggshell and exposing the other half to graded hyperoxia that doubled the oxygen gradient across the shell. Thus, the overall gas conductance was effectively normal but highly inhomogeneous. No differences were observed between the experimental and the control eggs in respiration, residual yolk or embryo growth rate from day 12 until day 20. The embryo was able to utilise the increase of oxygen in the environment to compensate for the loss of about half of its respiratory surface area under the wax. The results indicate that an even distribution of pores in the eggshell and uniformity of chorioallantoic perfusion are not required for successful avian development.

Animals↗
PubMed1998

The effect of hyperoxia on embryonic and organ mass in the developing chick embryo.

It is known that hyperoxia stimulates growth late in incubation when the chick embryo outgrows the O2 diffusion capacity. We wondered whether hyperoxia could have an effect in the early period prior to the stage where metabolism exceeds the oxygen diffusion capacity of the eggshell. For this we studied four groups of chicken eggs: control group (CG; n = 100) and three test groups (TGs) exposed during 48 h to 60% O2 on days 10, 14, and 18. In the CG, embryonic and organ mass (brain, heart, lungs, liver and intestine) were measured from day 10 until day 21 of incubation. In the TGs embryonic and organ mass were obtained from 24 h after the start of hyperoxia exposure until the end of incubation. In all TGs the most striking growth rate acceleration was observed in the liver and intestine, maximum growth rate accelerations were respectively, 19 and 42% in TG1, 43 and 173% in TG2 and 39% and 84 in TG3. In contrast, the brain was little affected by the hyperoxia exposure, the maximum growth rate acceleration was 14% in TG2. The results suggest that also in the middle of the incubation period O2 availability can be a limiting factor for growth, before metabolism exceeds the oxygen diffusion capacity of the eggshell.

Animals↗