The rate of oxygen consumption during embryogenesis of Lymnaea stagnalis (Gastropoda)

We studied the rate of oxygen consumption by the Lymnaea stagnalis embryos. The rate of oxygen consumption increased consistently during embryogenesis. The volume specific rate of oxygen consumption increased initially from the early cleavage stages until the gastrula stage and then decreased gradually to the eclosion of snails. There are three periods in embryogenesis of L. stagnalis, which differ in the coefficients of allometric dependence between the rate of oxygen consumption and volume of embryos: (1) early embryogenesis, when the increase in the rate of oxygen consumption is not accompanied by the growth of volume of the embryos; (2) larval period (trochophore and veliger stages; exponential coefficient k = 0.514), and (3) postlarval period (exponential coefficient k = 0.206).     

The rate of oxygen consumption during embryogenesis of Lymnaea stagnalis (Gastropoda)

We studied the rate of oxygen consumption by the Lymnaea stagnalis embryos. The rate of oxygen consumption increased consistently during embryogenesis. The volume specific rate of oxygen consumption increased initially from the early cleavage stages until the gastrula stage and then decreased gradually to the eclosion of snails. There are three periods in embryogenesis of L. stagnalis, which differ in the coefficients of allometric dependence between the rate of oxygen consumption and volume of embryos: (1) early embryogenesis, when the increase in the rate of oxygen consumption is not accompanied by the growth of volume of the embryos; (2) larval period (trochophore and veliger stages; exponential coefficient k = 0.514), and (3) postlarval period (exponential coefficient k = 0.206).     

Cardiac development in crayfish: ontogeny of cardiac physiology and aerobic metabolism in the red swamp crayfish Procambarus Clarkii

The cardiovascular system performs key physiological functions even as it develops and grows. The ontogeny of cardiac physiology was studied throughout embryonic and larval development in the red swamp crayfish Procambarus clarkii using videomicroscopic dimensional analysis. The heart begins to contract by day 13 of development (at 25°C, 20 kPa O₂). Prior to eclosion, heart rate (ƒH) decreases significantly. Previous data suggests that the decrease in cardiac parameters prior to hatching may be due to an oxygen limitation of the embryo. Throughout development, metabolizing mass and embryonic oxygen consumption primarily increased while egg surface area remains constant. The limited area for gas exchange of the egg membrane, in combination with the increasing oxygen demand of the embryo could result 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 (PO₂ =40 kPa O₂). The ƒH in late stage embryos increased significantly over control values when exposed to hyperoxic water suggesting that the suppression in cardiac function observed in late stage embryos is likely due to a limited oxygen supply.     

Individual growth of the great ramshorn snail <Emphasis Type="Italic">Planorbarius corneus</Emphasis> (Gastropoda,Planorbidae) embryos

Individual growth of the great ramshorn snail Planorbarius corneus has been studied by intravital video imaging. As has been observed, the types of growth change over the embryogenesis. The linear dimensions slightly but in a statistically significant manner decrease during the stages of cleavage to blastula. Starting from the stage of blastula to trochophore, the embryo diameter remains constant to commence increasing at the stage of middle trochophore. During the larval stages (trochophore and veliger), the growth is synchronous (in Dettlaffs, biological time units) for the embryos in both the same clutch and different clutches. The growth at that time is exponential but later desynchronizes in individual clutches. The embryos in eight clutches grew and developed slower and hatched later as compared with the remaining five egg clutches. An accelerated growth follows an asymptomatic pattern according to the von Bertalanffy equation. A retarded growth is describable with a linear equation. The observed differences are likely to be associated with the number of embryos in a clutch. All types of changes in the linear dimensions observed in the great ramshorn snail embryogenesis can be described with the same united equation.     

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.     

Quantification of embryo quality by respirometry

It is generally accepted that assessment of embryo metabolism, in particular oxygen consumption, may improve embryo selection by identifying the embryos with higher developmental competence. Several methods have been employed to measure embryonic oxygen consumption, but most of them were detrimental to subsequent embryo development. Recently, we have introduced the Nanorespirometer system, which is a non-invasive and highly sensitive technology developed for the individual measurement of embryonic respiration rates. This technology is able to perform single measurements at a fixed time or stage of embryonic development without adversely influencing embryo viability. Concomitantly, and based on the same principles, a second technology -- the Embryo Respirometer -- has been developed. The Embryo Respirometer allows the continuous measurement of individual respiration rates with simultaneous acquisition of digital images of each embryo, during the entire culture period (6-7 days). In this review, both technologies are described and their potential use as diagnostic tools for improving embryo selection in bovine and human following IVF treatments is discussed. Correlations between respiration rates of individual embryos and other parameters such as morphological quality, sex, stage of development, kinetics, diameter, expression of key metabolic genes and subsequent viability following embryo transfer are also examined. On the basis of the results obtained, it is postulated that assessment of embryonic respiration rates in association with other viability parameters allows for a more accurate embryo evaluation, both under clinical and research conditions.     

Dynamics of body mass and oxygen consumption in the ontogeny of the Spanish ribbed newt (<Emphasis Type="Italic">Pleurodeles waltl</Emphasis>): 2. Larval stage

The correlation between characteristics of growth and energy metabolism during the larval stage of development of the Spanish ribbed newt (Pleurodeles waltl) has been studied. During this period, its body mass is found to increase 140 times and the oxygen consumption rate, 77 times. The highest rate of specific body mass increase and oxygen consumption rate are noted in the early larval stage. Later, these characteristics decrease except for a brief period before completion of metamorphosis when the rate specific body mass increase rises. Comparison of the studied characteristics allows us to note a similar pattern in changes of the specific growth rate and the oxygen consumption rate during the premetamorphic development of the Spanish ribbed newt.     

Oxygen consumption and energy metabolism of the early mouse embryo

Oxygen consumption of preimplantation and early postimplantation mouse embryos has been measured using a novel noninvasive ultramicrofluorescence technique, based on an oil-soluble, nontoxic quaternary benzoid compound pyrene, whose fluorescence is quenched in the presence of oxygen. Pyruvate and glucose consumption, lactate production, and glycogen formation from glucose were also measured. Preimplantation mouse embryos of the strain CBA/Ca × C57BL/6 were cultured in groups of 10–30 in 2 μl of modified M2 medium containing 1 mmol l⁻¹ glucose, 0 mmol l⁻¹ lactate, and 0.33 mmol l⁻¹ pyruvate, for between 4–6 hr. Day 6.5 and 7.5 embryos were cultured singly in 40 μl M2 medium for between 2–3 hr. Oxygen consumption was detected at all stages of development, including, for the first time, in the early postimplantation embryo. Consumption remained relatively constant from zygote to morula stages before increasing in the blastocyst and day 6.5–7.5 stages. When expressed as QO₂ (μl/mg dry weight/hr), oxygen consumption was relatively constant from the one-cell to morula stages before increasing sharply at the blastocyst stage and declining to preblastocyst levels on days 6.5 and 7.5. Pyruvate was consumed during preimplantation stages, with glucose uptake undetectable until the blastocyst stage. Glucose was the main substrate consumed by the 6.5 and 7.5 day embryo. The proportions of glucose accounted for by lactate appearance were 81%, 86%, and 119% at blastocyst, day 6.5, and day 7.5 stages, respectively. The equivalent figures for glucose incorporated into glycogen were 10.36%, 0.21%, and 0.19%, respectively. The data are consistent with a switch from a metabolism dependent on aerobic respiration during early preimplantation stages to one dependent on both oxidative phosphorylation and aerobic glycolysis at the blastocyst stage, a pattern which is maintained on days 6.5 and 7.5. Our technique for measuring oxygen consumption may have diagnostic potential for selecting viable embryos for transfer following assisted conception techniques in man and domestic animals. © 1996 Wiley-Liss, Inc.     

Respiration of starfish oocytes during meiosis, fertilization, and artificial activation

Rates of oxygen consumption were measured in oocytes of the starfish Patiria miniata prior to and after the initiation of meiotic maturation in response to 1-methyladenine. No significant change in the rate of respiration was noted until after the completion of meiosis, at which point a two-fold increase in the rate of respiration was observed. The rate of oxygen consumption was also measured in response to fertilization and artificial activation with pronase. A transitory “burst” of oxygen consumption was noted in response to both stimuli. This “burst” is larger and of briefer duration in pronase-treated eggs. Possible interpretations of these phenomena are discussed.     

Deviation of the living system from the stationary state during oogenesis

Summary The changes in respiration and glycolysis of whole oocytes and homogenates of oocytes during oogenesis have been studied.The respiration rate of whole oocytes increases during oocyte growth and decreases during oocyte maturation. The respiration rate of homogenates also increases during oocyte growth and does not change during egg maturation. At all oogenesis stages the respiration rate of homogenates is higher than the respiration rate of whole oocytes.Respiration intensity increases during the small growth stage and decreases during the following stages of oogenesis. Respiration intensity of homogenates under optimal conditions changes in a similar way. Respiration intensity under physiological conditions diminishes during oogenesis from 70% at the small growth stage to 42% in unfertilised eggs.The rate of glycolysis in whole oocytes and homogenates of oocytes increases during the growth period of oocytes but does not change during egg maturation.Glycolysis intensity of the whole oocytes increases at the large growth stage—stage of cytoplasmic vacuolisation—and becomes less during the following stages. Glycolysis intensity in homogenates under optimal conditions is much higher than the glycolysis intensity of whole oocytes and it decreases slightly during oogenesis. The efficiency of glycolysis in oocytes under physiological conditions is very low. It increases from the stage of cytoplasmic vacuolisation (3.6%) to the stage at which vitellogenesis starts (20%) and diminishes at the following stages.The data obtained are considered in the light of the Prigogine and Wiame interpretation of a thermodynamic theory of development.     

Major muscle systems in the larval caenogastropod,Ilyanassa obsoleta,display different patterns of development

This study describes the anatomical and developmental aspects of muscular development from the early embryo to competent larval stage in the gastropod Ilyanassa obsoleta. Staining of F‐actin revealed differential spatial and temporal patterns of several muscles. In particular, two major muscles, the larval retractor and pedal retractor muscles originate independently and display distinct developmental patterns similar to observations in other gastropod species. Additionally, together with the larval retractor muscle, the accessory larval muscle developed in the embryo at the trochophore stage. Therefore, both these muscles develop prior to ontogenetic torsion. The pedal retractor muscle marked the most abundant growth in the mid veliger stage. Also during the middle stage, the metapodial retractor muscle and opercular retractor muscle grew concurrently with development of the foot. We show evidence that juvenile muscles, such as the buccal mass muscle and siphon muscle develop initially during the late veliger stage. Collectively, these findings substantiate that larval myogenesis involves a complex sequence of events that appear evolutionary conserved within the gastropods, and set the stage for future studies using this model species to address issues concerning the evolution and eventual fates of larval musculature in molluscs. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.     

Effects of biotic and abiotic factors on the oxygen content of green sea turtle nests during embryogenesis

Several biotic and abiotic factors can influence nest oxygen content during embryogenesis. Several of these factors were determined during each developmental stage of green sea turtle embryos on Wan-an Island, Penghu Archipelago, Taiwan. We examined oxygen content in 7 nests in 2007 and 11 in 2008. Oxygen in the adjacent sand, total and viable clutch sizes, air, sand and nest temperatures, and sand characters of each nest were also determined. Oxygen content was lower in late stages than in the early and middle stages. It was also lower in the middle layer than in the upper and bottom layers. Nest temperature showed opposite trends, reaching its maximum value in late stages of development. Nest oxygen content was influenced by fraction of viable eggs, total clutch sizes, sand temperatures, maximum nest temperature and maximum change in the nest temperature during incubation. Clutch size during embryogenesis was the most influential factor overall. However, the major influential factors were different for different developmental stages. In the first half of the incubation, the development rate was low, and the change in the nest oxygen content was influenced mainly by the clutch size. During the second half, the rapid embryonic development rate became the dominant factor, and hatchling activities caused even greater oxygen consumption during the last stage of development.     

Oxygen consumption by Nephrops norvegicus (L.), (Crustacea: Decapoda) in relationship with its moulting stage

The study of the rate of oxygen consumption by Nephrops norvegicius (L.) at each moulting stage has been carried out in closed respiration chambers, using an oxygen electrode to measure the oxygen concentration.Although the mean respiration rate seems to be low when compared with other decapod crustaceans of similar weight, the general trends of the variation of the oxygen consumption during the moulting cycle are similar to those described for other species of decapods. It should be noted that, during the process of the ecdysis, there is no oxygen consumption.     

Changes in the mitochondrial proteome of developing maize seed embryos

Mitochondria are required for seed development, but little information is available about their function and role during this process. We isolated the mitochondria from developing maize (Zea mays L. cv. Nongda 108) embryos and investigated the mitochondrial membrane integrity and respiration as well as the mitochondrial proteome using two proteomic methods, the two‐dimensional gel electrophoresis (2‐DE) and sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH). Mitochondrial membrane integrity and respiration were maintained at a high level up to 21 days after pollination (DAP) and decreased thereafter, while total mitochondrial number, cytochrome c oxidase activity and respiration per embryo exhibited a bell‐shaped change with peaks at 35–45 DAP. A total of 286 mitochondrial proteins changed in abundance during embryo development. During early stages of seed development (up to 21 DAP), proteins involved in energy production, basic metabolism, protein import and folding as well as removal of reactive oxygen species dominated, while during mid or late stages (35–70 DAP), some stress‐ and detoxification‐related proteins increased in abundance. Our study, for the first time, depicted a relatively comprehensive map of energy production by mitochondria during embryo development. The results revealed that mitochondria were very active during the early stages of maize embryo development, while at the late stages of development, the mitochondria became more quiescent, but well‐protected, presumably to ensure that the embryo passes through maturation, drying and long‐term storage. These results advance our understanding of seed development at the organelle level.     

Role of Respiration in the Germination Process of the Pathogenic Mold <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis>

Inhalation of resting conidia is usually the first step of a systemic infection caused by the opportunistic fungal pathogen Aspergillus fumigatus. In the lung, the inhaled spores encounter an environment that permits germination. However, the relative importance of certain environmental conditions for conidial activation and subsequent hyphae formation has so far not been analyzed in detail. In this study, we studied the role of oxygen during germination. We found that inhibitors of the respiratory chain were nearly as efficient in blocking germination as cycloheximide, an inhibitor of protein synthesis, which is already known to prevent germination of Aspergillus nidulans. We also found that A. fumigatus is unable to grow or germinate under anaerobic conditions, and using the fluorescent mitotracker dye we detected active mitochondria already at the stage of swollen conidia, which indicates that respiration is an early event during germination. In line with these data, we found that significant oxygen consumption was detectable early during germination, whereas no oxygen consumption was measurable in suspensions of resting conidia. In summary, the present study provides evidence that respiration is absolutely required for the germination of A. fumigatus conidia. Anela Taubitz and Bettina Bauer contributed equally.     

Cellular Basis of Neuralization of Induced Neurectoderm in Amphibian Embryogenesis: Changes of Cell Shape, Cell Size, and Cytodifferentiation of the Neurectoderm after Neural Induction

Cellular alterations of the neurectoderm after primary embryonic induction were examined by measuring several indices of shape, volume, and cytodifferentiation of the neurectodermal cells of Cynops embryos during gastrulation and early neurulation.
Results showed that cellular alterations occurs just after the 18 hr embryo stage (stage 13b). The thickness of the neurectoderm layer decreases like that of the epidermal ectoderm during early and middle gastrulation. After the 18 hr embryo stage, however, the neurectoderm thickens, mainly due to formation of columnar cells. Measurement of cell volume indicates that the neurectoderm of the early and middle gastrulae consists of a cell population of heterogeneous size. The heterogeneity diminishes sharply after the 18 hr embryo stage and the neural plate of the 36 hr embryo (stage 18) consists of cells of homogeneous size.
Stages before the 12 hr embryo (stage 12b) and after the 18 hr embryo (stage 13b) also showed differed in cell adhesion to the culture flask and in cytodifferentiating potency. Single cells dissociated from the neurectoderm of 18 hr embryos could adhere to the substratum and differentiate into both nerve-like cells and pigment cells. Both capacities increase during further development.
These results are discussed in relation to the neuralizing determination of neurectoderm after primary embryonic induction.     

杏花开放过程中超弱发光和ATP及活性氧含量的变化

以巴旦水杏 (Prunus armeniaca,Ameniacavulgaris Lam.)为试验材料,研究了杏花开放过程中花朵的超弱发光(ultraweak 1 uminescence,UWL)变化及其与ATP含量、呼吸速率、活性氧产生速率等的关系.结果表明,杏花开放的前半期(盛花期以前),超氧自由基产生速率与丙二醛含量变化平缓,UWL、ATP含量、呼吸速率及蛋白质含量呈上升趋势.盛花期过后,呼吸速率、蛋白质和ATP含量迅速降低,UWL强度缓慢下降,而超氧自由基的产生速率与丙二醛含量迅速升高.分析后认为,杏花的超微弱发光与以ATP含量为标志的能量水平密切相关,同时也受体内活性氧及其它物质代谢的影响;盛花期以前ATP可能是导致UWL升高的主要因素,盛花期过后,活性氧可能对花体的UWL变化起了部分的调节作用.     

Gene expression in early development of Lymnaea stagnalis

Pond snails (Lymnaea stagnalis) were irradiated with doses of 2760 r or 5520 r in order to induce lethal factors in the germ cells. The descendants of these irradiated snails were individually reared to adulthood, when by self-fertilization they produced egg masses. The occurrence of lethal factors was studied in these egg masses and the stage at which they manifested themselves was determined. No developmental disturbance was found during cleavage, but gastrulation was affected. This indicates that gastrulation in Lymnaea is controlled by the genome of the embryo. The stage at which most genes interfere with development is the early trochophore stage. At later stages the number of genes, which for the first time are active in development, declines. Since the late veliger stage was never affected, all genes essential for development, are apparently functioning before this stage.