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Tracing the evolutionary origin of the TFF-domain, an ancient motif at mucous surfaces.

Mammalian trefoil factor family (TFF)-domain peptides promote gastrointestinal protection, healing and cell migration and may act as tumour suppressors. TFF-like domains also constitute modules of composite proteins like mucin glycoproteins and zona pellucida proteins. Database searches with a modified, less stringent consensus sequence - C-x(5,6)-[ST]-x(3)-C-x(4,5)-C-C-[FYWH]-x(2, 24)-C-[FY] - revealed that ancestors of the TFF-domain arose before amphibian evolution. Eggshell proteins and a zona pellucida-like protein in teleost species, an epidermis-specific protein in a tunicate as well as an open reading frame in a nematode exhibited TFF-like motifs suggesting that they most likely originated in some multicellular organism.

Amino Acid Sequence↗

Interaction with avian cells and colonisation of specific pathogen free chicks by Shiga-toxin negative Escherichia coli O157:H7 (NCTC 12900).

The prevalence of Escherichia coli O157:H7 infection in birds is low but several deliberate inoculation studies show that poultry are readily and persistently infected by this organism indicating a possible threat to public health. The mechanisms of colonisation of poultry are not understood and the aim is to establish models to study the interaction of E. coli O157:H7, at the cellular and whole animal levels. A non-toxigenic E. coli O157:H7 (NCTC 12900) was used in adherence assays with an avian epithelial cell line (Div-1) and used to inoculate 1-day-old SPF chicks. In vitro, NCTC 12900 induced micro-colonies associated with cytoskeletal arrangements and pedestal formation with intimate bacterial attachment. In the 1-day-old SPF chick, a dose of 1 x 10(5) cfu resulted in rapid and extensive colonisation of the gastrointestinal tract and transient colonisation of the liver and spleen. The number of E. coli O157:H7 organisms attained approximately 10(8) cfu/ml caecal homogenate 24h after inoculation and approximately 10(7) cfu/ml caecal homogenate was still present at day 92. Faecal shedding persisted for 169 days, ceasing 9 days after the birds came into lay and 6% of eggs were contaminated on the eggshell. Histological analysis of tissue samples from birds dosed with 1x10(7) cfu gave evidence for E. coli O157:H7 NCTC 12900 induced micro-colonies on the caecal mucosa, although evidence for attaching effacing lesions was equivocal. These models may be suitable to study those factors of E. coli O157:H7 that mediate persistent colonisation in avian species.

Animals↗

Schistosoma mansoni: effect of recombinant tumor necrosis factor alpha on fecundity and [14C]-tyrosine uptake in females maintained in vitro.

It has been reported that recombinant tumor necrosis factor alpha (rTNF alpha) stimulates egg-laying in Schistosoma mansoni females. Because tyrosine requirement is increased in female undergoing sexual maturation in preparation for oogenesis and tyrosine is a major component of eggshell protein, we wanted to determine whether females treated with rTNF alpha would also incorporate more tyrosine. Adult females were first treated with 10, 20 or 40 ng/ml rTNF alpha for 1, 3 or 6 hr in RPMI 1640 containing 10% fetal calf serum (FCS). Another two groups of females were each exposed either to males or to male excretory-secretory (ES) products for 1 hr. They were then exposed to 20 microCi/ml-1 [14C]-tyrosine for 1 hr in RPMI 1640 containing 10% FCS. All females were incubated individually unless indicated otherwise. Females incorporated more tyrosine after exposure to males or their ES products. They incorporated significantly more tyrosine when treated with rTNF alpha for 1 hr; the increased uptake correlated with increasing amounts of rTNF alpha used. Although after a 3-hr treatment with 10 ng/ml rTNF alpha females incorporated slightly more tyrosine than controls, increasing amounts of rTNF alpha had an adverse effect. Females treated with rTNF alpha for 6 hr incorporated less tyrosine than controls and those treated for 1 hr. SDS-PAGE and fluorography did not reveal any differences in polypeptide profiles of untreated and rTNF alpha-treated females. These unexpected results led us to study the effect of rTNF alpha on fecundity in females. Contrary to the published report, we observed a sharp decline in egg-laying in females when exposed to increasing concentrations of rTNF alpha in vitro.

Animals↗

An extraosseous aneurysmal cyst in the pelvic cavity: report of a case.

A 51-year-old Japanese female was referred to us with a left lower quadrant pain and palpable mass. The CT and MRI study showed a 10x8x8 cm of well-circumscribed, multicystic mass adjacent to the left iliac bone. The mass was excised and consisted of multiple cysts containing bloody viscous material surrounded by thin-ring of eggshell-like tissue. The histological findings correlated with those of an aneurysmal bone cyst. This extraosseous case might be the first reported case observed in the pelvic cavity.

Bone Cysts, Aneurysmal↗

A huge osteoblastoma with aneurysmal bone cyst in skull base.

We reported a case of a huge osteoblastoma in the skull base of a young girl who had developed blindness in her right eye 1 year prior to examination. CT showed a large expansile bony lesion with eggshell margin over the skull base. MR imaging showed mixed solid and cystic parts with multiple fluid-fluid levels in the lesion. The optic chiasma and bilateral optic nerves were severely compressed by the tumor.

Adolescent↗

Evidence for increased hydroxylation of pyrrolidone amino acid residues in the cuticle of mature Onchocerca volvulus.

To determine whether morphologic changes are accompanied by variations in the biochemical and antigenic properties of the cuticle of Onchocerca volvulus during development, we isolated and compared the 2-mercaptoethanol soluble cuticular proteins and the insoluble cuticlin from the predominant life-cycle stages occurring in man. SDS-PAGE analysis, before and after digestion with collagenase from Achromobacter iophagus, revealed that the polypeptide composition of the 2-mercaptoethanol-solubilised extracts from adult males and nodular microfilariae are quite distinct and that these extracts contained predominantly collagen-like proteins. Demonstrated by immunoblotting with a hyper immune patient serum pool (n = 107), five strongly reactive antigens with apparent molecular weights of 126, 68, 43, 37 and 33 kDa were detected in the extracts from adult males, while at least eight prominent and several weakly reactive components were detected in the extracts from nodular microfilariae. The overall amino acid composition of the cuticular extracts from the various stages demonstrates that: (a) the cuticle of the adult male stage is rich in glycine, pyrrolidone amino acids, and acidic amino acids or their amides, (b) eggshells are particularly poor in proline but rich in serine residues (14.5%), (c) nodular microfilariae cuticular extracts are poor in proline but rich in valine (9.0%) and lysine (7.3%) and (d) hydroxyproline and hydroxylysine are present in the cuticle of adults but absent in the juvenile life-cycle stages (nodular microfilariae and eggs). This study firstly, indicates that the composition of the cuticle of O. volvulus may thus, be quite distinct from one parasite stage to another and secondly, that the maturation of the parasite in the human host may be accompanied by the extensive hydroxylation of prolyl residues and to a lesser extent of lysyl residues in the predominantly collagen-like cuticular proteins.

Amino Acids↗

Command and control: regulatory pathways controlling invasive behavior of the border cells.

The invasiveness of cancer cells resembles the normal behavior of cells that migrate into surrounding tissues during development. For example, the border cells in the Drosophila ovary undergo a partial epithelial to mesenchymal transition and invade the neighboring cluster of germline cells, migrating to the oocyte border. Once there, they provide patterning information to the oocyte and produce an eggshell specialization known as the micropyle. Border cell migration has been subjected to extensive genetic analyses using a variety of screening approaches. Recent findings demonstrate that conversion of the border cells from a stationary group of epithelial cells to invasive cells requires integration of the activities of at least two transcriptional regulatory pathways. One such pathway requires the slbo gene, which encodes Drosophila C/EBP, a basic region/leucine zipper transcriptional activator that is required for elevated expression of a number of downstream targets, including DE-cadherin and focal adhesion kinase (FAK). An independent pathway requires the activity of the ecdysone receptor and a recently identified co-activator for the ecdysone receptor known as Taiman (abbreviated TAI, pronounced ti-maan', meaning too slow). Ecdysone is produced in the Drosophila ovary in response to adequate nutrition and is required for progression of oogenesis through stage 9, when border cell migration occurs. Border cells mutant for tai accumulate abnormally high levels of adhesion complexes at their surfaces, which may account for their inability to migrate. Thus border cell migration requires a differentiation program mediated by the C/EBP pathway, which is required for elevated expression of a number of proteins required for motility. In addition, migration requires a hormonal signal that relays information regarding nutritional status and appears to be required for regulation of the proper localization of some of the C/EBP targets. These findings suggest that steroid hormones can regulate cell motility relatively directly, independent of the effects on proliferation. This may contribute to the metastatic effects of steroid hormones on certain cancers and the inhibition of metastasis by steroid hormone antagonists such as tamoxifen.

Animals↗

Regulation of osteopontin gene expression during egg shell formation in the laying hen by mechanical strain.

The aim of this study is to evaluate the regulation of the osteopontin (OPN) gene expression by non-hormonal stimuli, such as calcium flux and mechanical strain during the daily egg cycle in the oviduct of the laying hen. After the egg enters the eggshell gland (ESG), the OPN gene is expressed by the epithelium cells in two waves: first by the basal cells and only then by the apical cells of the epithelium. A reduction in OPN gene expression was observed 1 h prior to laying. The calbindin gene, which marks the onset of calcification, was found to be expressed in the glandular epithelium starting 2 h after OPN gene expression. In addition, the formation of soft shells was accompanied by a reduction in calbindin, but not in OPN, gene expression. The application of a mechanical strain comparable to that induced by an egg led to induction of OPN gene expression at a normally quiescent phase in the cyclical expression of this gene. The induction of the gene was time- and strain-dependent and temporally similar to that induced by the entry of the egg into the ESG. In contrast, the calbindin gene was not affected by mechanical strain. The ESG of the laying hen provides a system to study the effect of a mechanical strain on matrix protein production in vivo, in a relevant physiological setting. The finding suggests that, in contrast to calbindin, OPN gene expression is not regulated by calcium flux but rather by the mechanical strain imposed by the resident egg.

Animals↗

Is beta-pleated sheet the molecular conformation which dictates formation of helicoidal cuticle?

Over 100 sequences for cuticular proteins are now available, but there have been no formal analyses of how these sequences might contribute to the helicoidal architecture of cuticle or to the interaction of these proteins with chitin. A secondary structure prediction scheme (Hamodrakas, S.J., 1988. A protein secondary structure prediction scheme for the IBM PC and compatibles. CABIOS 4, 473-477) that combines six different algorithms predicting alpha-helix, beta-strands and beta-turn/loops/coil has been used to predict the secondary structure of chorion proteins and experimental confirmation has established its utility (Hamodrakas, S.J., 1992. Molecular architecture of helicoidal proteinaceous eggshells. In: Case, S.T. (Ed.), Results and Problems in Cell Differentiation, Vol. 19, Berlin-Heidelberg, Springer Verlag, pp. 116-186 and references therein). We have used this same scheme with eight cuticular protein sequences associated with hard cuticles and nineteen from soft cuticles. Secondary structure predictions were restricted to a conserved 68 amino acid region that begins with a preponderance of hydrophilic residues and ends with a 33 amino acid consensus region, first identified by Rebers and Riddiford (Rebers, J.F., Riddiford, L.M., 1988. Structure and expression of a Manduca sexta larval cuticle gene homologous to Drosophila cuticle genes. J. Mol. Biol. 203, 411-423). Both classes of sequences showed a preponderance of beta-pleated sheet, with four distinct strands in the proteins from 'hard' cuticles and three from 'soft'. In both cases, tyrosine and phenylalanine were found on one face within a sheet, an optimal location for interaction with chitin. We propose that this beta-sheet dictates formation of helicoidal cuticle.

Amino Acid Sequence↗

Avian reproductive anatomy, physiology and endocrinology.

Although many environmental cues influence reproductive activity, the seasonal breeder responds most strongly to long day length. THE MALE BIRD: Testicular interstitial cells secrete testosterone, which influences reproductive behavior such as territorial aggression and song. Other changes observed in seasonal breeders include testicular hypertrophy and enlargement of the ductus deferens and seminal glomus. THE FEMALE BIRD: Early changes associated with rising estrogen levels in the hen include osteomyelosclerosis and hypercalcemia. Ovulation is then induced by LH, which is followed by eggshell calcification, which is under the control of progesterone. Sources of calcium for shell production include intestinal absorption from the diet, renal control of calcium levels, and mobilization of bone calcium stores. During oviposition, PGF2 alpha and vasotocin stimulate powerful uterine contractions [32] in the presence of calcium. Incubation is associated with falling LH levels and rising prolactin levels. If the hen actually enters reproductive quiescence at this time, then molt will follow. Molt is associated with the total regression of the reproductive tract.

Animals↗

Evolution of placentation among squamate reptiles: recent research and future directions.

Squamate reptiles are uniquely suited to study of evolution of reproductive mode and pattern of embryonic nutrition. Viviparous species have evolved from oviparous ancestors on numerous occasions, patterns of nutritional provision to embryos range widely from lecithotrophy, at one end of a continuum, to placentotrophy at the other, and structure and function of the maternal-embryonic relationship is highly constrained resulting in parallel evolutionary trajectories among taxa. Embryos of oviparous species primarily receive nourishment from yolk, but also mobilize a significant quantity of calcium from the eggshell. Most viviparous species also are predominantly lecithotrophic, yet all viviparous species are placentotrophic to some degree. Similarities in embryonic development and nutritional pattern between oviparous species and most viviparous species suggest that the pattern of nutrition of oviparous squamates is an exaptation for the evolution of viviparity and that placentotrophy and viviparity evolve concomitantly. The few species of squamates that rely substantially on placentotrophy have structural modifications of the interface between the embryo and mother that are interpreted as adaptations to enhance nutritional exchange. Recent studies have extended understanding of the diversity of embryonic nutrition and placental structure and have resulted in hypotheses for transitions in the evolution of placentotrophy, yet data are available for few species. Indirect tests of these hypotheses, by comparison of structural-functional relationships among clades in which viviparity has evolved, awaits further study of the reproductive biology of squamates.

Animals↗

Utilisation of lipids, protein, ions and energy during embryonic development of Australian oviparous skinks in the genus Lampropholis.

The contents of eggs and neonates of the Australian skinks, Lampropholis guichenoti and L. delicata, are described and compared to allow interpretation of nutrient utilisation by the developing embryo. Even though the females are the same size, L. guichenoti lay smaller clutches of larger eggs (egg contents=41.6+/-1.2 mg dry mass) than L. delicata (26.6+/-2.8 mg). The energy density is the same for eggs (30.5+/-0.9 J/g ash-free dry mass for L. guichenoti and 29.9+/-1.1 J/mg for L. delicata) and neonates (22.5+/-1.3 J/mg for L. guichenoti and 23.5+/-0.4 J/mg for L. delicata) between species. The amount of nitrogen (protein) in neonates is only slightly lower than that in eggs, whereas there is a large and significant decline in total lipids. Thus, like some other skinks, protein is a source of metabolic energy during embryogenesis, although not as important as lipid. Triacylglycerol is the major lipid component of the eggs (80% of total lipid), with phospholipid forming only approximately 10% of the total lipid. The fatty acid profile of the phospholipid is distinguished by a high proportion of arachidonic acid (8%), a significant proportion of eicosapentaenoic acid (2-4%) and a relatively low proportion of docosahexaenoic acid (2-3%) compared to chickens. Eggs of both species have remarkably low concentrations of free cholesterol compared to other amniote eggs (0.7% for L. guichenoti and 1.3% for L. delicata). The loss of lipid during embryonic development is almost entirely due to the selective utilisation of yolk triacylglycerol, presumably for energy. By contrast, the amount of phospholipid recovered from the neonates was the same as that originally in the eggs. Moreover, significantly more total cholesterol was present in the neonates than in the eggs, suggesting that biosynthesis of additional cholesterol occurred during development. The phospholipids of the neonates contain higher proportions of arachidonic (11-12%) and docosahexaenoic (8%) acids than the phospholipids of the eggs. Eicosapentaenoic acid is less prevalent in phospholipids in neonates than in eggs. Neonates of both species contain significantly more calcium than the fresh egg contents (L. guichenoti, eggs 0.303+/-0.051 mg, neonates 0.641+/-0.047 mg; L. delicata, eggs 0.187+/-0.013 mg, neonates 0.435+/-0.033 mg), presumably as a result of resorption of calcium from the eggshell. Interestingly, there is also significantly more sodium in neonates than in the contents of fresh eggs (L. guichenoti, eggs 0.094+/-0.010 mg, neonates 0.184+/-0.011 mg; L. delicata, eggs 0.084+/-0.011 mg, neonates 0.151+/-0.010 mg). There is no significant difference in the content of potassium and magnesium in eggs and neonates of either species. Although the fresh eggs of L. delicata have a significantly higher sodium concentration than L. guichenoti, there is no difference in the concentrations of calcium, magnesium, potassium or sodium in the neonates of the two species.

Animals↗

Development of the pulmonary surfactant system in non-mammalian amniotes.

Pulmonary surfactant (PS) is a complex mixture of phospholipids, neutral lipids and proteins that lines the inner surface of the lung. Here, it modulates surface tension thereby increasing lung compliance and preventing the transudation of fluid. In mammals, the PS system develops towards the end of gestation, characterized by an increase in the saturation of phospholipids in lung washings and the appearance of surfactant proteins in amniotic fluid. Birth, the transition from in utero to the external environment, is a rapid process. At this time, the PS system is important in opening and clearing the lung of fluid in order to initiate pulmonary ventilation. In oviparous vertebrates, escape from an egg can be a long and exhausting process. The young commence pulmonary ventilation and hatching by 'pipping' through the eggshell, where they remain for some time, presumably clearing their lungs. This paper relates changes in the development of the pulmonary surfactant system within the non-mammalian amniotes in response to birth strategy, lung morphology and phylogeny in order to determine the conservatism of this developmental process. Total phospholipid (PL), disaturated phospholipid (DSP) and cholesterol (Chol) were quantified from lung washings of embryonic and hatchling chickens, bearded dragons (oviparous), sleepy lizards (viviparous), snapping turtles and green sea turtles throughout the final stages of incubation and gestation. In all cases, the pattern of development of the pulmonary surfactant lipids was consistent with that of mammals. PL and DSP increased throughout the latter stages of development and Chol was differentially regulated from the PLs. Maximal secretion of both PL and DSP occurred at 'pipping' in oviparous reptiles, coincident with the onset of airbreathing. Similarly, the amount of DSP relative to total PL was maximal immediately after the initiation of airbreathing in chickens. The relative timing of the appearance of the lipids differed between groups. In the oviparous lizard, surfactant lipids were released over a relatively shorter time than that of the sleepy lizard, turtles, birds and mammals. Thus, despite temporal differences and vastly different lung morphologies, birth strategies and phylogenies, the overall development and maturation of the PS system is highly conserved amongst the amniotes.

Animals↗

Gas exchange and energy metabolism of the ostrich (Struthio camelus) embryo.

We measured oxygen consumption (V(O(2))) and carbon dioxide emission (V(CO(2))) rates, air-cell gas partial pressures of oxygen (P(A)O(2)) and CO(2) (P(A)CO(2)), eggshell water vapour conductance and energy content of the ostrich (Struthio camelus) egg, 'true hatchling' and residual yolk, and calculated RQ and total oxygen consumption (V(O(2)tot)) for ostrich eggs incubated at 36.5 degrees C and 25% relative humidity. The V(O(2)) pattern showed a drop of approximately 5% before internal pipping. V(O(2)) just prior to internal pipping agrees with allometric calculations. Despite the higher incubation temperature compared to other studies, and the resultant shorter incubation duration (42 days), V(O(2)tot) (91.7 l kg(-1)) was similar to a previously reported value. RQ values during the second half of incubation (approx. 0.68) were lower than expected for lipid catabolism. Prior to internal pipping, P(A)O(2) and P(A)CO(2) were 98 and 48.3 torr (13.1 and 6.4 kPa), respectively. The growth pattern of the ostrich embryo is different from the typical precocial pattern, showing a time delay in the rapid growth phase. As a result, the lowered overall energy expenditure for tissue maintenance, as compared to other species, is reflected in the low yolk utilization and high residual yolk fraction of the whole hatchling dry mass. These could also result from the relatively short incubation period of the ostrich egg, thereby evading desiccation by excess water loss.

Animals↗

Cardiac rhythms in prenatal and perinatal emu embryos.

Emu eggs weigh approximately 600 g and have an incubation duration (ID) of approximately 50 days. The egg mass is approximately 10-fold heavier than the chicken egg and the ID is approximately 2.5-fold longer. Daily changes in mean heart rate (MHR) of emu embryos were previously determined, but further measurement was needed to investigate the species-specific behavior of cardiac rhythm for comparison with other species. In the present study, we continuously measured the electrocardiogram of emu embryos while maintaining adequate gas exchange through the eggshell and determined instantaneous heart rate (IHR) during the last 2-7 days of incubation until hatching or death. The MHR over 1-min intervals was calculated from IHR data in order to present continuous developmental patterns of heart rate (HR) in a single graph and 24-h recordings of HR in a single panel, showing the HR trend over a prolonged period. However, neither circadian nor ultradian rhythms of HR were shown in these figures or by power spectrum analysis. The IHR distinctively fluctuated and the fluctuations were mainly comprised of three patterns of irregular HR accelerations in embryos that hatched. Respiratory sinus arrhythmia also occurred in perinatal embryos. During the final stages of the perinatal period, short-term, repeated, large accelerations of IHR appeared, which signaled imminent hatching and has been reported for chick embryos. IHR fluctuations in embryos that failed to hatch tended to become inactive towards death.

Animals↗

Development of respiratory rhythms in perinatal chick embryos.

In chick embryos, gas exchange takes place via the chorioallantoic membrane (CAM) and the lungs at approximately 1 day prior to hatching. The present study was designed to elucidate the development of respiratory rhythms in the chick embryo during the whole pipping (perinatal) period with a condenser-microphone measuring system. The microphone was hermetically attached on the eggshell over the air cell on day 18 of incubation. It first detected a cardiogenic signal (i.e. acoustocardiogram), and then beak clapping and breathing signals (acoustorespirogram, ARG). The first signals of lung ventilation appeared intermittently and irregularly approximately once per 5 s among the clapping signals after the embryo penetrated its beak into the air cell (internal pipping, IP). The respiratory rhythm then developed irregularly, with a subsequent more regular rate. The envelope pattern of breathing from the onset of IP through external pipping (EP) to hatching was constructed by a specially devised procedure, which eliminated external and internal noises. The envelope patterns indicated that the IP, EP and whole perinatal periods of 10 embryos were 14.1+/-6.4 (S.D.), 13.6+/-4.0 and 27.6+/-5.4 h, respectively. In addition, they also indicated the period of embryonic hatching activity (i.e. climax) which was 48+/-19 min. The development of respiratory rhythm was also shown by the instantaneous respiratory rate (IRR) which was designated as an inverse value of two adjacent ARG waves.

Animals↗

Energy and nutrient utilisation by embryonic reptiles.

Most reptiles are oviparous, with the developing embryos relying on the contents of the yolk to sustain development until hatching (lecithotrophy). The yolk is composed primarily of lipid and protein, which act as an energy source and the essential components to build embryonic tissue. Nevertheless, yolk and the resulting embryos contain many other nutrients, including inorganic ions, vitamins, carotenoids, water and hormones. Apart from water and oxygen, which may be taken up by eggs, and some inorganic ions that can come from the eggshell or even from outside the egg, everything required by the embryo must be in the egg when it is laid. Approximately 20% of squamate reptiles are viviparous, exhibiting a variety of placental complexities. Species with complex placentae have reduced yolk volumes, with the mother augmenting embryonic nutrition by provision across the placenta (placentotrophy). Despite assumed advantages of placentotrophy, only 5 out of approximately 100 lineages of viviparous squamates exhibit substantial placentotrophy. This paper reviews available and recent information on the yolk contents of a variety of squamate reptiles to ask the question, how are nutrients transported from the yolk to the embryo or across the placenta? Although, current available data suggest that, in broad terms, yolk is taken up by embryos without discrimination of the nutrients, there are some apparent exceptions, including the very long chain polyunsaturated fatty acids. In addition, fundamental differences in the patterns of energy utilisation in lizards and snakes suggest fundamental differences in lipid profiles in these taxa, which appear to reflect the differences between placentotrophic and lecithotrophic viviparous lizards.

Animals↗

Cardiac rhythms of late pre-pipped and pipped chick embryos exposed to altered oxygen environments.

During the final stages of embryonic development in chickens, diffusive gas exchange through the chorioallantoic membrane (CAM) is progressively replaced by pulmonary respiration that begins with internal pipping (IP) of the CAM. Late chick embryos going through the transition from CAM respiration to pulmonary respiration were exposed to hyperoxic (100% O(2)) and hypoxic (10% O(2)/N(2)) environments for 2-h and the responses of baseline heart rate (HR), and HR fluctuation patterns were investigated. 16- and 18-day-old (referred to as 18-d) embryos and 20-d externally pipped (EP) embryos were examined as pre-pipped embryos and pipped embryos, respectively. 19-d embryos were divided into two groups: embryos that had not yet internally pipped (Pre-IP embryos) and embryos that had internally pipped (IP embryos). IP was identified by detecting the breathing signal with a condenser microphone attached hermetically on the eggshell (i.e. acoustorespirogram) on day 19 of incubation. In the hyperoxic environment, HR baseline of pre-pipped embryos remained unchanged and that of pipped embryos was depressed. In the hypoxic environment, HR baseline of 16-d pre-pipped embryos was depressed and that of pipped (IP and EP) embryos elevated. These different responses in pipped embryos might be partially attributed to increased cholinergic input from the vagus nerve in hyperoxia and increased adrenergic response in hypoxia. While hyperoxia did not induce marked modification of instantaneous heart rate (IHR) fluctuation patterns, hypoxia tended to augment transient decelerations of IHR in late pre-pipped embryos and markedly depressed HR fluctuations in pipped embryos.

Allantois↗