Physiological development of the embryonic and larval crayfish heart

The cardiovascular system is the first system to become functional in a developing animal and must perform key physiological functions even as it develops and grows. The ontogeny of cardiac physiology was studied throughout embryonic and larval developmental stages in the red swamp crayfish Procambarus clarkii using videomicroscopic dimensional analysis. The heart begins to contract by day 13 of development (at 25 degrees C, 20 kPa O(2)). Cardiac output is primarily regulated by changes in heart rate because stroke volume remains relatively constant throughout embryogenesis. Prior to eclosion, heart rate and cardiac output decreased significantly. Previous data suggest that the decrease in cardiac parameters prior to hatching may be due to an oxygen limitation to the embryo. Throughout development, metabolizing mass and embryonic oxygen consumption increased, while egg surface area remained constant. The surface area of the egg membrane is a constraint on gas exchange; this limitation, in combination with the increasing oxygen demand of the embryo, results in an inadequate diffusive supply of oxygen to developing tissues. To determine if the decrease in cardiac function was the result of an internal hypoxia experienced during late embryonic development, early and late-stage embryos were exposed to hyperoxic water (PO(2) = 40 kPa O(2)). Heart rate in late-stage embryos exposed to hyperoxic water increased significantly over control values, which suggests that the suppression in cardiac function observed in late-stage embryos is due to a limited oxygen supply.     

Effects of environmental oxygen on development and respiration of Australian lungfish (<Emphasis Type="Italic">Neoceratodus forsteri</Emphasis>) embryos

The effects of oxygen partial pressure ( P_\textO2 P_{{{\text{O}}_{2} }} ) on development and respiration were investigated in the eggs of the Australian lungfish, Neoceratodus forsteri. At 20°C, embryonic survival and development was optimal at 15 and 20.9 kPa. Development was slowed at 5 and 10 kPa and embryos did not survive 2 kPa. At lower P_\textO2 P_{{{\text{O}}_{2} }} , the rate of oxygen consumption also decreased. Embryos responded to hypoxia by hatching at an earlier age and stage of development, and hatching wet and dry gut-free masses were reduced. The role of oxygen conductance ( G_\textO2 G_{{{\text{O}}_{2} }} ) in gas exchange was also examined under selected environmental P_\textO2 P_{{{\text{O}}_{2} }} and temperatures. The breakdown of the vitelline membrane changed capsule geometry, allowed water to be absorbed into the perivitelline space and increased capsule G_\textO2 G_{{{\text{O}}_{2} }} . This occurred at embryonic stage 32 under all treatments and was largely independent of both P_\textO2 P_{{{\text{O}}_{2} }} and temperature (15, 20 and 25°C), demonstrating that capsule G_\textO2 G_{{{\text{O}}_{2} }} cannot adaptively respond to altered environmental conditions. The membrane breakdown increased capsule diffusive G_\textO2 G_{{{\text{O}}_{2} }} and stabilised perivitelline P_\textO2 P_{{{\text{O}}_{2} }} , but reduced the convective G_\textO2 G_{{{\text{O}}_{2} }} of the perivitelline fluid, as the large perivitelline volume and inadequate convective current resulted in a P_\textO2 P_{{{\text{O}}_{2} }} gradient within the egg prior to hatch.     

Respiratory and cardiovascular responses to acute hypoxia and hyperoxia in internally pipped chicken embryos

During the first day of hatching, the developing chicken embryo internally pips the air cell and relies on both the lungs and chorioallantoic membrane (CAM) for gas exchange. Our objective in this study was to examine respiratory and cardiovascular responses to acute changes in oxygen at the air cell or the rest of the egg during internal pipping. We measured lung (V·_O2lung) and CAM (V·_O2CAM) oxygen consumption independently before and after 60 min exposure to combinations of hypoxia, hyperoxia, and normoxia to the air cell and the remaining egg. Significant changes in V·_O2total were only observed with combined egg and air cell hypoxia (decreased V·_O2total) or egg hyperoxia and air cell hypoxia (increased V·_O2total). In response to the different O₂ treatments, a change in V·_O2lung was compensated by an inverse change in V·_O2CAM of similar magnitude. To test for the underlying mechanism, we focused on ventilation and cardiovascular responses during hypoxic and hyperoxic air cell exposure. Ventilation frequency and minute ventilation (V_E) were unaffected by changes in air cell O₂, but tidal volume (V_T) increased during hypoxia. Both V_T and V_E decreased significantly in response to decreased P_CO2. The right-to-left shunt of blood away from the lungs increased significantly during hypoxic air cell exposure and decreased significantly during hyperoxic exposure. These results demonstrate the internally pipped embryo's ability to control the site of gas exchange by means of altering blood flow between the lungs and CAM.     

Serotonergic sensory‐motor neurons mediate a behavioral response to hypoxia in pond snail embryos

Oxygen (O₂) is one of the most important environmental factors that affects both physiological processes and development of aerobic animals, yet little is known about the neural mechanism of O₂ sensing and adaptive responses to low O₂ (hypoxia) during development. In the pond snail, Helisoma trivolvis, the first embryonic neurons (ENC1s) to develop are a pair of serotonergic sensory‐motor cells that regulate a cilia‐driven rotational behavior. Here, we report that the ENC1‐ciliary cell circuit mediates an adaptive behavioral response to hypoxia. Exposure of egg masses to hypoxia elicited a dose‐dependent and reversible acceleration of embryonic rotation that mixed capsular fluid, thereby facilitating O₂ diffusion to the embryo. The O₂ partial pressures (Po₂) for threshold, half‐maximal, and maximal rotational response were 60, 28, and 13 mm Hg, respectively. During hypoxia, embryos relocated to the periphery of the egg masses where higher Po₂ levels occurred. Furthermore, intermittent hypoxia treatments induced a sensitization of the rotational response. In isolated ciliary cells, ciliary beating was unaffected by hypoxia, suggesting that in the embryo, O₂ sensing occurs upstream of the motile cilia. The rotational response of embryos to hypoxia was attenuated by application of the serotonin receptor antagonist, mianserin, correlated to the development of ENC1‐ciliary cell circuit, and abolished by laser‐ablation of ENC1s. Together, these data suggest that ENC1s are unique oxygen sensors that may provide a good single cell model for the examination of mechanistic, developmental, and evolutionary aspects of O₂ sensing. © 2002 Wiley Periodicals, Inc. J Neurobiol 52: 73–83, 2002     

Isolation and identification of substances inducing formation of tracheary elements in cultured carrot-root slices

Data are presented on the cytokinin status of seeds and seed components, at different stages of development in Phaseolus coccineus L., as determined with the soybean callus growth bioassay: A change in cytokinin types according to developmental stage occurred: from biologically very active less polar types (zeatin=Z) at early stages to more polar types (zeatin glucoside=Z₉G and zeatin riboside=Zr), with relatively low biological activity, at intermediate and late stages of seed development: When cytokinins were analyzed separately in embryos (embryo proper) and suspensors at two embryonic stages: heart-shaped (A) and middle cotyledonary embryos (stage B) respectively, it was found that: i) at stage A, the suspensor showed cytokinin activity at the level of Z, 2iPA (2-isopentenyladenosine) and Zr, whereas more polar cytokinins (Z₉G, Zr) were present in the embryo; ii) at stage B, when the embryo seems to become autonomous for cytokinin supply, there was a relative abundance of active cytokinins (Z, 2iPA) in the embryo to which Z₉G activity in the suspensor corresponded. It is concluded that the suspensor plays an essential role in embryogenesis by acting as a hormone source to the early embryo.Abbreviations GA gibberellic acid - 2iPA 2-isopentenyladenosine - Stage A heart-shaped embryo - siage B middle cotyledonary embryo - Z zeatin - Z₉G zeatin glucoside - Zr Zeatin riboside     

Heart rate responses to altered ambient oxygen in early (days 3–9) chick embryos in the intact egg

Normal heart rate (HR), and the HR responses to hypoxia and hyperoxia during early heart development in chick embyros have not been studied in detail, particularly in undisturbed embryos within the intact egg. HR was measured in day 3–9 chick embryos at 38 °C using relatively noninvasive impedance cardiography. Embryos were exposed to air (control) and to hypoxic (10% O₂) or hyperoxic (100% O₂) gas for a 2-h or 4-h period, during which HR was continually monitored. Control (normoxic) HR increased from about 150 beats per min (bpm) on day 3 to about 240 bpm on days 7–9. HR in very early embryos showed a variety of moderate responses to hypoxia (all survived), but as development progressed beyond day 6, hypoxic exposure induced a profound bradycardia that frequently terminated in death before the end of the measurement period. In contrast to the marked developmental changes in hypoxic sensitivity, HR showed little response to hyperoxia throughout development, suggesting no “hypoxic drive” to HR. We speculate that hypoxia has little effect early in development because of the embryo's small absolute O₂ demand, but as the embryo grows, hypoxia represents a progressively more severe perturbation. Although general trends were identified, there was considerable variation in both HR and HR responses to ambient O₂ changes between individuals of the same developmental stage. Accepted: 16 December 1998     

δ18O‐derived incubation temperatures of oviraptorosaur eggs

In order to determine the incubation temperature of eggs laid by non‐avian dinosaurs, we analysed the oxygen isotope compositions of both eggshell carbonate (δ¹⁸O_c) and embryo bone phosphate (δ¹⁸O_p) from seven oviraptorosaur eggs with preserved in ovo embryo bones. These eggs come from the Upper Cretaceous Nanxiong Formation of Jiangxi Province, China. Oviraptorosaur theropods were selected because of their known brooding behaviour as evidenced by preserved adult specimens fossilized in brooding posture on their clutch. Incubation temperature of these embryos was estimated based on the following considerations: eggshell δ¹⁸O_c value reflects the oxygen isotope composition of egg water fluid; embryo bones precipitate from the same egg fluid; and oxygen isotope fractionation between phosphate and water is controlled by the egg temperature. A time‐dependent model predicting the δ¹⁸O_p evolution of the embryo skeleton during incubation as a function of egg temperature was built, and measured δ¹⁸O_c and δ¹⁸O_p values used as boundary conditions. According to the model outputs, oviraptorosaurs incubated their eggs within a 35–40°C range, similar to extant birds and compatible with the known active brooding behaviour of these theropod dinosaurs. Provided that both eggshell and embryo bones preserved their original oxygen isotope compositions, this method could be extended to investigate some reproductive traits of other extinct groups of oviparous amniotes.     

Water chemistry in the microenvironment of rainbow trout Oncorhynchus mykiss embryos is affected by development,the egg capsule and crowding

The hypothesis tested was that embryonic metabolism affects the water chemistry in the boundary layer. In addition, embryo crowding would further compound the metabolic effect on the water chemistry in the boundary layer. As development progressed, the magnitude of the boundary layer gradients for O₂ and pH, but not for NH, increased. The presence of the egg capsule hindered the diffusion of O₂ into and H⁺ and NH out of the embryo. The magnitude of the O₂, pH and NH boundary layer gradient was significantly increased when embryos were surrounded by either sham embryos or live embryos. The majority of this crowding effect on embryo boundary layers was due to changes in water flow rather than due to metabolism directly. These results clearly show that the microenvironment adjacent to the developing rainbow trout Oncorhynchus mykiss embryo becomes more stagnant as development progresses in the presence of the egg capsule and is further intensified with embryo crowding.     

Der embryonale Sauerstoffverbrauch des Buntspechts (Dendrocopos major): Einflu? von Entwicklungsmodus und Bebrütungszeit

Zusammenfassung Der Sauerstoffverbauch von zwei Eiern des Buntspechtes (Dendrocopos major) wurde während der letzten sechs Tage der Embryogenese mit Hilfe eines Durchflußrespirometers gemessen. Der zeitliche Verlauf des embryonalen Stoffwechsels folgt annähernd einer Exponentialfunktion mit Ausnahme einer siebenstündigen Plateauphase am 9. Bebrütungstag. Während der gesamten Embryogenese setzt der Buntspecht 0.9 kJ × g^–1 Frischeimasse um. Dies entspricht nur etwa dem halben Umsatz durchschnittlicher Vogelembryonen (rund 2 kJ × g^–1). Als Ursache dafür wird die kurze Bebrütungszeit des Buntspechts (10 Tage) diskutiert.

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Embryonic oxygen consumption in the Great Spotted Woodpecker (Dendrocopos major) — influence of developmental mode and incubation period

Oxygen consumption of two eggs of the Great Spotted Woodpecker was measured with a two-channel oxygen analyzer using an open flow system. The embryogeny of oxygen consumption nearly follows an exponential equation with the exception of a 7-hour plateau phase on day 9 of incubation. During this period the embryos consume ca. 4 ml O₂ × min^–1. For a given fresh egg mass the eggs of woodpeckers have unusually short incubation periods (10 days measured vs. 17 days expected). Thus a comparison of the embryonic metabolism with that of species having long incubation periods but similar egg sizes shows the influence of a fast embryonic development on avian enbryonic energetics. The total embryonic metabolism of the Great Spotted Woodpecker amounts to 0.9 kJx g^–1 fresh egg mass which is half as much as the value for most other birds (ca. 2 kJ × g^–1).

     

Study of the late embryogenesis of Daphnia (Anomopoda, ‘Cladocera’, Branchiopoda) and a comparison of development in Anomopoda and Ctenopoda

The embryonic development of Daphnia galeata and D. hyalina (`Cladocera', Anomopoda, Daphniidae) has been investigated by observing living embryos removed from female brood pouches. The sequence of morphological changes was analysed, as was the time at which the activity of certain organs began. The timing of these events at 22 °C is documented for both species.These data were compared with similar information, previously obtained for two representatives of the Ctenopoda (Kotov & Boikova, 1998). The sequence of events is basically similar in the two groups during early and late phases of their development, but the time of shedding of the embryonic membranes is different in the Anomopoda and Ctenopoda. The ctenopod embryo hatching from the second egg membrane is covered by the third membrane, which will be cast some hours later. The anomopod embryo hatches from the second egg membrane approximately simultaneously with the shedding of the third membrane, and it is covered already by the fourth membrane after the shedding of the second egg membrane.Earlier (Kotov & Boikova, 1998), we determined four embryonic instars in the course of the development of the Ctenopoda. Two of them are passed within the egg membranes, the next two instars occur after the shedding the egg membranes within the mother's brood pouch. However, in anomopods, one of the latter (the third) occurs within the second egg membrane, one is incorporated into the egg. Thus, the development of the Anomopoda is more embryonized in comparison with that of the Ctenopoda.     

Role of the left aortic arch and blood flows in embryonic American alligator (<Emphasis Type="Italic">Alligator mississippiensis</Emphasis>)

All embryonic and fetal amniotes possess a ductus(i) arteriosus(i) that allows blood to bypass the pulmonary circulation and the non-functional lungs. The central hemodynamic of embryonic reptiles are unique, given the additional systemic aorta that allows pulmonary circulatory bypass, the left aorta (LAo). The LAo exits in the right ventricle or ‘pulmonary side’ of reptilian hearts in both embryos and adults, but its functional significance in ovo is unknown. This study investigated the role of the LAo in embryonic American alligators by surgically occluding the LAo and measuring oxygen consumption and, in addition, measured hemodynamic responses to hypoxia in embryonic alligators. We measured systemic cardiac output and primary chorioallantoic membrane (CAM) artery blood flow for normoxic and hypoxic-incubated (10% O₂) American alligator embryos (Alligator mississippiensis). Chronic blood flow (1–124 h) in the primary CAM artery for hypoxic-incubated embryos (92 ± 26 ml min⁻¹ kg⁻¹) was elevated when compared with normoxic-incubated embryos (29 ± 14 ml min⁻¹ kg⁻¹, N = 6; P = 0.039). For hypoxic-incubated embryos, acute LAo blood flow (49.6 ± 24.4 ml min⁻¹ kg⁻¹) was equivalent to the combined flow of the three systemic great vessels that arise from the left ventricle, the right aorta, common carotid and subclavian arteries (43.6 ± 21.5 ml min⁻¹ kg⁻¹, N = 5). Similarly, for normoxic-incubated embryos, LAo blood flow (27.3 ± 6.6 ml min⁻¹ kg⁻¹) did not statistically differ from the other three vessels (18.4 ± 4.9 ml min⁻¹ kg⁻¹, N = 5). This study contains the first direct test of LAo function and the first measurements of blood flow in an embryonic reptile. These data support the hypotheses that embryonic alligators utilize the LAo to divert a significant amount of right ventricular blood into the systemic circulation, and that CAM blood flow increases following chronic hypoxic conditions. However, surgical occlusion of the LAo did not affect egg [(V)\dot]_\textO2, \dot{V}_{{\text{O}}_{2}}, supporting the hypothesis that the LAo of reptiles is not critical to maintain in ovo oxygen consumption.     

Simulating in ovulo osmotic potentials and O<Subscript>2</Subscript> tensions normalize growth and pigmentation of immature cotton embryos

Oxygen tensions and osmotic potentials are important physiological factors of plant growth and development. To optimize these variables for cotton (Gossypium hirsutum L.) embryo culture, we quantified dissolved O₂ (dO₂) tensions, osmotic potentials, and pH at several locations in cotton ovules during embryony. Clark O₂ microelectrodes were micromanipulated into intact ovules at an angle lateral to the developing embryo, and dO₂ tensions were determined in integuments, nucelli and embryos. Ovular osmotic potentials and pH were determined from extracted ovule sap using vapor pressure osmometers and pH microelectrodes. Dissolved O₂ tensions near or in embryos decreased from 104 mmol m⁻³ at 5 days post-anthesis (DPA) to 83 mmol m⁻³ at 18 DPA. Osmotic potentials of ovule sap decreased from −0.70 megapascals (MPa) at 2 DPA to −1.12 MPa at 8 DPA but then increased to −0.84 MPa by 17 DPA. Ovule sap pH at 5–17 DPA varied inconsistently and ranged from 5.4 to 6.5. Based on these results, a factorial experiment with two osmotic and three O₂ treatments was designed. Immature embryos of cotton cultivar HS-26 were randomly assigned to the treatment combinations and cultured for 33 days. Oxygen treatments did not affect embryo growth, and there were no differences among treatments with regard to percentage of embryos that progressed to a more advanced stage of embryo development. However, cotyledons of embryos grown without osmotic adjustment were abnormally large, and embryos exposed to these treatments were abnormally brown. Browning was less severe for embryos exposed to low O₂ tensions. Growth and pigmentation were most normal for embryos simultaneously exposed to O₂ tensions and osmotic potentials that best simulated the observed in ovulo conditions.     

Compatible invasion of a phylogenetically distant host embryo by a hymenopteran parasitoid embryo

Embryonic invasion into the tissue of genetically different organisms has been known only in mother-embryo interactions of viviparous organisms. Hence, embryonic invasions have been thought to occur only within the same or closely related species. For endoparasitic Hymenoptera, which are oviposited in their host egg but complete their development in the later stages, entry into the host embryo is essential. To date, the entry of these parasitoids is known to be accomplished by either egg deposition directly into the embryo or by the newly hatched larva boring into the embryo. However, Copidosoma floridanum is a polyembryonic parasitoid whose development is characterized by a prolonged embryonic stage, and which lacks a larval form during its host embryogenesis. We have analyzed the behavior and fate of C. floridanum embryos co-cultured with their host embryo in vitro. Here, we show that the morula-stage embryo of C. floridanum actively invades the host embryo. Histological analyses have demonstrated that C. floridanum embryonic invasion is associated with adherent junction to host cells rather than causing an obvious wound on the host cells. These findings provide a novel case of embryonic invasion into a phylogenetically distant host embryo, ensuring cellular compatibility with host tissues.     

Mouse Embryonic Development in a Serum-free Whole Embryo Culture System

Mid-gestation stage mouse embryos were cultured utilizing a serum-free culture medium prepared from commercially available stem cell media supplements in an oxygenated rolling bottle culture system. Mouse embryos at E10.5 were carefully isolated from the uterus with intact yolk sac and in a process involving precise surgical maneuver the embryos were gently exteriorized from the yolk sac while maintaining the vascular continuity of the embryo with the yolk sac. Compared to embryos prepared with intact yolk sac or with the yolk sac removed, these embryos exhibited superior survival rate and developmental progression when cultured under similar conditions. We show that these mouse embryos, when cultured in a defined medium in an atmosphere of 95% O₂ / 5% CO₂ in a rolling bottle culture apparatus at 37 °​C for 16-40 hr, exhibit morphological growth and development comparable to the embryos developing in utero. We believe this method will be useful for investigators needing to utilize whole embryo culture to study signaling interactions important in embryonic organogenesis.     

Late embryo-defective mutants of Arabidopsis

The embryo-defective (emb) mutants of Arabidopsis constitute a large and diverse group of mutants disrupted in a broad range of embryonic processes, including morphogonesis, cell differentiation, and maturation programs. This report describes a subset of these mutants, the late embryo defectives, which develop beyond the globular stage of embryogenesis but fail to complete normal morphogenesis. A representative sample of 12 late mutants was chosen for this study, patterns of morphogenesis were characterized, the germination potential of mutant seeds was investigated, and additional mutant alleles within the collection were identified. Morphological defects in mutant embryos became apparent during the heart stage of development, when embryos normally begin the rapid cell division and expansion required for the completion of morphogenesis. Despite their morphological abnormalities, mutant embryos often germinated from dry seed, demonstrating that genetic programs required for the establishment of desiccation tolerance remained intact. Mutant seedlings displayed a wide range of developmental abnormalities, including altered morphology, lack of pigmentation, dwarfism, and disorganized vegetative growth. One late mutant was found to be allelic to an early embryo defective that arrests at the globular stage. These results suggest that a number of late EMB genes encode basic cellular and metabolic functions needed for cell division, enlargement, and embryonic growth. The rapid growth and metabolic changes that occur at the heart stage may present a barrier to normal development in the late mutants, resulting in altered embryo morphology and other developmental defects. It is proposed that many Arabidopsis mutants with abnormal embryo and seedling morphology are not defective in the regulation of pattern formation or morphogenesis, but rather in fundamental physiological and cellular processes required for the completion of normal growth and development. © 1995 Wiley-Liss, Inc.     

Gas exchange through the jelly capsule of the terrestrial eggs of the frog,Pseudophryne bibroni

Summary The terrestrial eggs of the Australian frog,Pseudophryne bibroni, increase in diameter during development by absorbing water from the substrate into the perivitelline space. This decreases the thickness of the jelly capsule and increases its effective surface area, both effects joining to increase the O₂ conductance of the capsule in parallel with the rate of O₂ consumption. PerivitellineP _{O 2}, calculated from capsule morphology and metabolic rate, and confirmed with micro-O₂-electrodes, remains high and constant (17–19 kPa) throughout the 39 day incubation period at 12°C. The increase in conductance is essential for adequate oxygenation of late embryos. Eggs incubated on substrates of 0 and –25 kPa matric tension (bracketing field conditions) take up disparate amounts of water but show similar conductance.     

Zygote shrinkage and subsequent development in some Hibiscus hybrids

Summary Early embryonic development was compared in self-fertilized embryos of the diploid species, Hibiscus costatus, and triploid hybrid embryos, H. costatus-aculeatus and H. costatus-furcellatus, the paternal parent in both hybrids being tetraploid. The self-fertilized zygotes shrank to 50% of the volume of the unfertilized egg. These young embryos showed marked polarity. There was a concentration of cytoplasm in the apical cells and large vacuoles in the basal cells. There was also a polar distribution of organelles within the embryo as a whole which probably reflected initial differentiation. In comparison, hybrid zygotes shrank only about 20% of their original volume but started division at about the same time as selffertilized zygotes. There appeared to be no polarization and little proliferation of the cytoplasm in the hybrids. Large vacuoles remained prominent throughout the hybrid embryos, while organelles were few in the scant cytoplasm and no polarization of these was evident. These highly divergent hybrid embryos had become necrotic and aborted by the time the normal, self-fertilized embryos had reached the late globular stage. This altered developmental sequence of the hybrids suggests that shrinkage and rearrangement of the zygote cytoplasm is essential for normal embryonic differentiation.     

Gas exchange and blood gases of Puna teal (Anas versicolor puna) embryos in the Peruvian Andes

Oxygen consumption, air cell gases, hematology, blood gases and pH of Puna teal (Anas versicolor puna) embryos were measured at the altitude at which the eggs were laid (4150 m) in the Peruvian Andes. In contrast to the metabolic depression described by other studies on avian embryos incubated above 3700 m, O₂ consumption of Puna teal embryos was higher than even that of some lowland avian embryos at equivalent body masses. Air cell O₂ tensions dropped from about 80 toor in eggs with small embryos to about 45 toor in eggs containing a 14-g embryo; simultaneously air cell CO₂ tension rose from virtually negligible amounts to around 26 torr. Arterial and venous O₂ tensions (32–38 and 10–12 toor, respectively, in 12- to 14-g embryos) were lower than described previously in similarly-sized lowland wild avian embryos or chicken embryos incubated in shells with restricted gas exchange. The difference between air cell and arterial O₂ tensions dropped significantly during incubation to a minimum of 11 torr, the lowest value recorded in any avian egg. Blood pH (mean 7.49) did not vary significantly during incubation. Hemoglobin concentration and hematocrits rose steadily throughout incubation to 11.5 g · 100 ml^-1 and 39.9%, respectively, in 14-g embryos.Abbreviations PO₂ partial pressure gradient of O₂ - BM body mass - D diffusion coefficient - G gas conductance (cm³·s^-1·torr^-1) - conductance to water vapor - IP internal pipping of embryos - P _ACO₂ partial pressure of carbon dioxide in air cell - P _AO₂ partial pressure of oxygen in air cell - P _aCO₂ partial pressure of carbon dioxide in arterial blood - P _aCO₂ partial pressure of oxygen in arteries - P _H barometric pressure (torr) - PCO₂ partial pressure of carbon dioxide - P _IO₂ partial pressure in ambiant air - PO₂ partial pressure of oxygen - P _VCO₂ venous carbon dioxide partial pressure - P _VO₂ mixed venous oxygen partial pressure - SE standard error - VO ₂ oxygen consumption     

Abiotic influences on embryo growth: statoliths as experimental tools in the squid early life history

Statolith size and growth was used to determine the influence of abiotic factors on the growth of Loligo vulgaris and Sepioteuthis australis embryos. Recently spawned egg masses collected from the field were incubated in the laboratory under different levels of light intensity, photoperiod, or short periods of low salinity (30‰). Double tetracycline staining was used to follow statolith growth. In L. vulgaris constant light conditions produced significantly slower growth in the embryonic statoliths and embryos held at summer photoperiod had slower statolith growth than those held at winter photoperiods. However once they hatched out there was no evidence that photoperiod affected statolith growth. After hatching, in all photoperiods statolith growth rates decreased in comparison with late embryonic rates. In S. australis embryos, differences between the high and medium light intensities for summer and intermediate photoperiods were found, suggesting that under summer incubation temperature, longer daylengths at medium light intensity favoured higher statolith growth for this species. In comparison to controls, slower statolith growth in S. australis embryos due to low salinity only occurred when exposed for 72 h. Comparison with previous studies indicates that temperature seems to be the main abiotic factor influencing statolith growth during early stages, however, interactions among all abiotic factors needs to be determined as well as the unknown influence of other isolated factors, e.g., oxygen concentration within the egg mass.