174. ATMOSPHERIC OXYGEN ALTERS THE EMBRYONIC METABOLOME AS QUANTIFIED BY CARBOHYDRATE UPTAKE AND AMINO ACID UTILISATION

2010 ◽  
Vol 22 (9) ◽  
pp. 92
Author(s):  
P. L. Wale ◽  
D. K. Gardner
Keyword(s):  
Amino Acid ◽  
Atmospheric Oxygen ◽  
Oxygen Toxicity ◽  
Glucose Consumption ◽  
Blastocyst Stage ◽  
Quadrupole Mass ◽  
F1 Hybrid ◽  
Amino Acid Utilization ◽  
Hybrid Mice ◽  
Amino Acid Utilisation

Oxygen regulates embryo development at both the cleavage and post compaction stages. In this study we investigated the effects of atmospheric oxygen during the preimplantation stages, on the post-compaction embryonic metabolome through quantification of glucose consumption and amino acid utilization. Zygotes obtained from F1 hybrid mice (C57BLxCBA) were randomly allocated to either 5 or 20% oxygen. In the first experiment, following the first 48 h of culture, embryos were cultured individually in 1 μL drops of modified G2 medium (0.5 mM glucose) and moved to fresh drops of medium every 24 h. The glucose concentration in the spent media samples, including controls containing no embryo, was determined by microfluorimetry. In the second experiment, embryos which had developed to the early blastocyst stage after 72 h were cultured for a further 24 h in groups of 10 in 2 μL drops of G2. Analysis of amino acid utilization was performed using liquid chromatography-triple quadrupole mass spectrometry. Glucose consumption by embryos cultured in 5% oxygen was significantly greater on day 4 and day 5 (4.89 ± 0.29 and 6.13 ± 0.41 pmol/embryo/h) compared to embryos cultured in 20% oxygen (2.59 ± 0.40 and 5.09 ± 0.28 pmol/embryo/h; P < 0.05). In contrast amino acid utilisation by embryos cultured in 5% oxygen was significantly less than embryos cultured in 20% oxygen (P < 0.05). The data generated will help to determine the aetiology of oxygen toxicity to the preimplantation embryo. Higher glucose utilisation by embryos in 5% oxygen is consistent with their improved development. Conversely, the increased utilisation of amino acids by blastocysts in 20% oxygen may reflect an adaptation to increased oxidative stress as a result of culture in a non-physiological oxygen concentration. This study demonstrates that atmospheric oxygen during the preimplantation period perturbs the embryonic metabolome which results in a compensatory increase in amino acid utilisation.

The roles of pyruvate, lactate and glucose during preimplantation development of embryos from F1 hybrid mice in vitro

Development ◽  
1991 ◽  
Vol 112 (1) ◽  
pp. 99-105 ◽  
Author(s):  
J.J. Brown ◽  
D.G. Whittingham
Keyword(s):  
Inbred Mouse ◽  
Blastocyst Stage ◽  
Preimplantation Development ◽  
F1 Hybrids ◽  
Mouse Strains ◽  
F1 Hybrid ◽  
Morula Stage ◽  
Lactate And Pyruvate ◽  
Hybrid Mice

Embryos of certain inbred mouse strains, and their F1 hybrids, are able to develop from the 1-cell to blastocyst stage in simple chemically defined media containing lactate (L), pyruvate (P) and glucose (G). The individual roles of these substrates in supporting complete preimplantation development in vitro was examined with 1-cell F2 embryos from B6CBF1 hybrid mice. Embryos collected between 26 and 27 h post hCG were cultured in medium containing L, P, LP or LPG. After 50 h in culture, the proportions developing to the morula stage were 1%, 83%, 94% and 100%, respectively. In combination, lactate and pyruvate appeared to act synergistically and both the rate and level of development to the morula stage were unaffected by the absence of glucose. After a further 46 h in culture, only the embryos grown in the presence of glucose developed into blastocysts. In LP medium, embryos arrested at the compacted morula stage late on day 3 of development. As culture continued in the absence of glucose, embryos decompacted (approximately 82 h post hCG) and subsequently degenerated. Exposure to medium containing glucose for the first, second or third 24 h period in culture was sufficient to support the morula-to-blastocyst transition. Glucose still supported this transition when embryos were transferred to LPG medium 3 h after the completion of compaction (76 h post hCG), but was ineffective 6 h later (82 h post hCG) once decompaction had commenced. We conclude that lactate and pyruvate together are able to support normal development of 1-cell F2 embryos to the morula stage in vitro, but that glucose is an essential component of the culture medium for development to the blastocyst stage.

scholarly journals Metabolic Activity of Human Embryos after Thawing Differs in Atmosphere with Different Oxygen Concentrations

2020 ◽  
Vol 9 (8) ◽  
pp. 2609
Author(s):  
Michal Ješeta ◽  
Andrea Celá ◽  
Jana Žáková ◽  
Aleš Mádr ◽  
Igor Crha ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Oxygen Concentration ◽  
Atmospheric Oxygen ◽  
Blastocyst Stage ◽  
Human Embryos ◽  
Total Amino Acid ◽  
Oxygen Levels ◽  
Amino Acid Turnover ◽  
Cultivation Environment

The vitrification of human embryos is more and more frequently being utilized as a method of assisted reproduction. For this technique, gentle treatment of the embryos after thawing is crucial. In this study, the balance of amino acids released to/consumed from the cultivation media surrounding the warmed embryos was observed in the context of a cultivation environment, which was with the atmospheric oxygen concentration ≈20% or with a regulated oxygen level—hysiological (5%). It is the first time that total amino acid turnover in human embryos after their freezing at post compaction stages has been evaluated. During this study, progressive embryos (developed to blastocyst stage) and stagnant embryos (without developmental progression) were analyzed. It was observed that the embryos cultivated in conditions of physiological oxygen levels (5% oxygen) showed a significantly lower consumption of amino acids from the cultivation media. Progressively developing embryos also had significantly lower total amino acid turnovers (consumption and production of amino acids) when cultured in conditions with physiological oxygen levels. Based on these results it seems that a cultivation environment with a reduced oxygen concentration decreases the risk of degenerative changes in the embryos after thawing. Therefore, the cultivation of thawed embryos in an environment with physiological oxygen levels may preclude embryonal stagnation, and can support the further development of human embryos after their thawing.

scholarly journals IDH1 deficiency attenuates gluconeogenesis in mouse liver by impairing amino acid utilization

2016 ◽  
Vol 114 (2) ◽  
pp. 292-297 ◽  
Author(s):  
Jing Ye ◽  
Yu Gu ◽  
Feng Zhang ◽  
Yuanlin Zhao ◽  
Yuan Yuan ◽  
...  
Keyword(s):  
Amino Acid ◽  
Hepg2 Cells ◽  
Mouse Liver ◽  
High Protein Diet ◽  
Glucose Consumption ◽  
Null Mouse ◽  
Isocitrate Dehydrogenase 1 ◽  
Amino Acid Utilization ◽  
Body Weights

Although the enzymatic activity of isocitrate dehydrogenase 1 (IDH1) was defined decades ago, its functions in vivo are not yet fully understood. Cytosolic IDH1 converts isocitrate to α-ketoglutarate (α-KG), a key metabolite regulating nitrogen homeostasis in catabolic pathways. It was thought that IDH1 might enhance lipid biosynthesis in liver or adipose tissue by generating NADPH, but we show here that lipid contents are relatively unchanged in both IDH1-null mouse liver and IDH1-deficient HepG2 cells generated using the CRISPR-Cas9 system. Instead, we found that IDH1 is critical for liver amino acid (AA) utilization. Body weights of IDH1-null mice fed a high-protein diet (HPD) were abnormally low. After prolonged fasting, IDH1-null mice exhibited decreased blood glucose but elevated blood alanine and glycine compared with wild-type (WT) controls. Similarly, in IDH1-deficient HepG2 cells, glucose consumption was increased, but alanine utilization and levels of intracellular α-KG and glutamate were reduced. In IDH1-deficient primary hepatocytes, gluconeogenesis as well as production of ammonia and urea were decreased. In IDH1-deficient whole livers, expression levels of genes involved in AA metabolism were reduced, whereas those involved in gluconeogenesis were up-regulated. Thus, IDH1 is critical for AA utilization in vivo and its deficiency attenuates gluconeogenesis primarily by impairing α-KG–dependent transamination of glucogenic AAs such as alanine.

scholarly journals Distinct profiles of human embryonic stem cell metabolism and mitochondria identified by oxygen

Reproduction ◽  
2015 ◽  
Vol 150 (4) ◽  
pp. 367-382 ◽  
Author(s):  
Jarmon G Lees ◽  
Joy Rathjen ◽  
John R Sheedy ◽  
David K Gardner ◽  
Alexandra J Harvey
Keyword(s):  
Amino Acid ◽  
Mitochondrial Function ◽  
Cell Function ◽  
Atmospheric Oxygen ◽  
Cell Metabolism ◽  
Embryonic Stem ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Mitochondrial Activity ◽  
Hes Cells

Oxygen is a powerful regulator of cell function and embryonic development. It has previously been determined that oxygen regulates human embryonic stem (hES) cell glycolytic and amino acid metabolism, but the effects on mitochondria are as yet unknown. Two hES cell lines (MEL1, MEL2) were analyzed to determine the role of 5% (physiological) and 20% (atmospheric) oxygen in regulating mitochondrial activity. In response to extended physiological oxygen culture, MEL2 hES cells displayed reduced mtDNA content, mitochondrial mass and expression of metabolic genesTFAM,NRF1,PPARaandMT-ND4. Furthermore, MEL2 hES cell glucose consumption, lactate production and amino acid turnover were elevated under physiological oxygen. In stark contrast, MEL1 hES cell amino acid and carbohydrate use and mitochondrial function were relatively unaltered in response to oxygen. Furthermore, differentiation kinetics were delayed in the MEL1 hES cell line following BMP4 treatment. Here we report the first incidence of metabolic dysfunction in a hES cell population, defined as a failure to respond to oxygen concentration through the modulation of metabolism, demonstrating that hES cells can be perturbed during culture despite exhibiting the defining characteristics of pluripotent cells. Collectively, these data reveal a central role for oxygen in the regulation of hES cell metabolism and mitochondrial function, whereby physiological oxygen promotes glucose flux and suppresses mitochondrial biogenesis and gene expression.

scholarly journals METABOLIC CONNECTION BETWEEN PROLINE AND GLYCINE IN THE AMINO ACID UTILIZATION OF TORULOPSIS UTILIS

1947 ◽  
Vol 169 (3) ◽  
pp. 759-760
Author(s):  
G. Ehrensvärd ◽  
E. Sperber ◽  
E. Saluste ◽  
L. Reio ◽  
R. Stjernholm
Keyword(s):  
Amino Acid ◽  
Amino Acid Utilization
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