scholarly journals The Effects of Storage in vitro on Functions, Transcriptome, Proteome, and Oxidation Resistance of Giant Grouper Sperm

2021 ◽  
Vol 8 ◽  
Author(s):  
Yang Yang ◽  
Tong Wang ◽  
Sen Yang ◽  
Xi Wu ◽  
Wenhua Huang ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Oxidative Phosphorylation ◽  
Oxidation Resistance ◽  
Cold Storage ◽  
Prolonged Storage ◽  
Negative Effects ◽  
Normal Functions ◽  
Oxidative Phosphorylation Pathway ◽  
The Absolute

Asynchrony of sexual maturity is a huge limitation in the reproduction of grouper sperm. Cold storage of sperm is an effective method to solve the problem of asynchronization. However, sperms gradually lose their activity with the prolonged storage time in vitro. In order to explore causes, the effects of cold storage on transcriptome, proteome and oxidation resistance of giant grouper sperm were analyzed. Firstly, the absolute RNA quantity and consistent transcripts existed in each spermatozoon were estimated. With the prolonged storage, the RNA quantity gradually decreased both in the cytoplasm and in the mitochondria of the spermatozoon. The decreased transcripts were mainly enriched with energy metabolism and stress response. Similar to RNAs, the absolute protein quantity was also significantly decreased during the storage of sperm. Decreased proteins were mainly enriched with the oxidative phosphorylation pathway. Proteins involved in the oxidative phosphorylation showed a faster degradation rate compared to the average total protein. In addition, the oxidation resistance and adenosine triphosphate (ATP) contents showed a significant decrease in the sperm during storage in vitro. These results implied that damages of transcriptome, proteome, and oxidation resistance have negative effects on the normal functions of sperm, especially their energy metabolism. The present study provides essential foundation for improving the storage of sperm in vitro.

scholarly journals Dynamics of Energy Metabolism in Carbon Starvation-Induced Fruitlet Abscission in Litchi

Horticulturae ◽  
2021 ◽  
Vol 7 (12) ◽  
pp. 576
Author(s):  
Qian Wu ◽  
Xingshuai Ma ◽  
Qingxin Chen ◽  
Ye Yuan ◽  
Huicong Wang ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Oxidative Phosphorylation ◽  
Citrate Synthase ◽  
Tca Cycle ◽  
Carbon Starvation ◽  
Fruit Abscission ◽  
Oxidative Phosphorylation Pathway ◽  
Glycolysis Pathway ◽  
Liquid Chromatography Tandem Mass ◽  
Fruitlet Abscission

Fruit abscission is triggered by multiple changes in endogenous components of the fruit, including energy metabolism. However, it is still unknown how the core energy metabolism pathways are modified during fruit abscission. Here, we investigated the relationship between carbon starvation-induced fruitlet abscission and energy metabolism changes in litchi. The fruitlet abscission of litchi ‘Feizixiao’ was induced sharply by girdling plus defoliation (GPD), a carbon stress treatment. Using liquid chromatography tandem mass spectrometry (LC-MS/MS) targeted metabolomics analysis, we identified a total of 21 metabolites involved in glycolysis, TCA cycle and oxidative phosphorylation pathways. Among them, the content of most metabolites in glycolysis pathways and TCA cycles was reduced, and the activity of corresponding metabolic enzymes such as ATP-dependent phosphofructokinase (ATP-PFK), pyruvate kinase (PK), citrate synthase (CS), succinate thiokinase (SAT), and NAD-dependent malate dehydrogenase (NAD-MDH) was decreased. Consistently, we further showed that the expression of the relative genes (LcPFK2, LcPK2, LcPK4, LcCS1, LcCS2, LcSAT, LcMDH1 and LcMDH2) was also significantly down-regulated. In contrast, the level of ATP, an important metabolite in the oxidative phosphorylation pathway, was elevated in parallel with both higher activity of H+-ATPase and the increased expression level of LcH+-ATPase1. In conclusion, our findings suggest that carbon starvation can induce fruitlet abscission in litchi probably by energy depletion that mediated through both the suppression of the glycolysis pathway and TCA cycle and the enhancement of the oxidative phosphorylation pathway.

scholarly journals In vitro methylation of bovine papillomavirus alters its ability to transform mouse cells.

1986 ◽  
Vol 6 (8) ◽  
pp. 2910-2915 ◽  
Author(s):  
B A Christy ◽  
G A Scangos
Keyword(s):  
Escherichia Coli ◽  
Bovine Papillomavirus ◽  
Negative Effects ◽  
Absolute Level ◽  
Mouse Cells ◽  
The Absolute ◽  
In Cells

Bovine papillomavirus (BPV) was methylated in vitro at either the 29 HpaII sites, the 27 HhaI sites, or both. Methylation of the HpaII sites reduced transformation by the virus two- to sixfold, while methylation at HhaI sites increased transformation two- to fourfold. DNA methylated at both HpaII and HhaI sites did not differ detectably from unmethylated DNA in its efficiency of transformation. These results indicate that specific methylation sites, rather than the absolute level of methylated cytosine residues, are important in determining the effects on transformation and that the negative effects of methylation at some sites can be compensated for by methylation at other sites. BPV molecules in cells transformed by methylated BPV DNA contained little or no methylation, indicating that the pattern of methylation was not faithfully retained in these extrachromosomally replicating molecules. Methylation at the HpaII sites (but not the HhaI sites) in the cloned BPV plasmid or in pBR322 also inhibited transformation of the plasmids into Escherichia coli HB101 cells.

scholarly journals Proliferation impairs cell viability via energy depletion

2020 ◽  
Author(s):  
Pierre Galichon ◽  
Morgane Lannoy ◽  
Li Li ◽  
Sophie Vandermeersch ◽  
David Legouis ◽  
...  
Keyword(s):  
Oxidative Phosphorylation ◽  
Cell Survival ◽  
Cell Lines ◽  
Renal Outcome ◽  
Energy Status ◽  
Transcriptomic Data ◽  
Intracellular Atp ◽  
Expression Of Genes ◽  
Oxidative Phosphorylation Pathway

ABSTRACTIntroductionProliferation is essential to the development of all living organisms and to the replacement of dead cells in injured organs. Proliferation is tightly regulated, and loss of proliferation control is a hallmark of cancer. Thus, cell proliferation and cell survival are closely interconnected to contribute to organ homeostasis or tumor development. Given the central role of energy homeostasis in cell survival, and the fact that proliferation increases negentropy, we hypothesized that proliferation might affect the intracellular ATP/ADP ratio, a robust readout of the cellular energy status.MethodsWe analyzed transcriptomic data and cytotoxicity assessment from tumor cell lines challenged with a panel of chemotherapies. We then analyzed the effect of proliferation on the viability and on the intracellular ATP/ADP ratio of epithelial cell lines challenged with toxic or energetic stresses. Finally, we studied transcriptomic data from both tumors and injured or recovering kidneys, and computed indexes for proliferation, and nuclear vs mitochondrially encoded oxidative phosphorylation genes.ResultsHere we found that proliferation is associated with decreased survival after toxic or energetic stresses in both cancer and epithelial cells. In vitro, we found that ATP/ADP ratio was tightly regulated throughout the cell cycle, and that proliferation was instrumental to an overall decrease in intracellular ATP/ADP ratio. In vivo, we found that the expression of genes of the oxidative phosphorylation pathway (OXPHOS) was correlated with proliferation in cancer. In injured kidneys, proliferation was also associated with increased expression of genes of the oxidative phosphorylation pathway encoded in the nucleus, but mitochondrially-encoded genes were strongly decreased, suggesting the coexistence of a passive mitochondrial injury and an adaptative nuclear response with opposite effects on OXPHOS. Increased proliferation and decreased expression of mitochondrially-encoded genes of the oxidative phosphorylation pathway were associated with a poor renal outcome. In sum, we show that proliferation is an energy demanding process impairing the cellular ability to cope with a toxic or ischemic injury.

Retinal pH Reflects Retinal Energy Metabolism in the Day and Night

2004 ◽  
Vol 91 (6) ◽  
pp. 2404-2412 ◽  
Author(s):  
Andrey V. Dmitriev ◽  
Stuart C. Mangel
Keyword(s):  
Energy Metabolism ◽  
Oxidative Phosphorylation ◽  
Energy Production ◽  
Ph Gradient ◽  
Living Tissue ◽  
Selective Suppression ◽  
Subjective Night ◽  
Circadian Time ◽  
The Brain

The extracellular pH of living tissue in the retina and elsewhere in the brain is lower than the pH of the surrounding milieu. We have shown that the pH gradient between the in vitro retina and the superfusion solution is regulated by a circadian (24-h) clock so that it is smaller in the subjective day than in the subjective night. We show here that the circadian changes in retinal pH result from a clock-mediated change in the generation of H+ that accompanies energy production. To demonstrate this, we suppressed energy metabolism and recorded the resultant reduction in the pH difference between the retina and superfusate. The magnitude of the reduction in the pH gradient correlated with the extent of energy metabolism suppression. We also examined whether the circadian-induced increase in acid production during the subjective night results from an increase in energy metabolism or from the selective activation of glycolysis compared with oxidative phosphorylation. We found that the selective suppression of either oxidative phosphorylation or glycolysis had almost identical effects on the dynamics and extent of H+ production during the subjective day and night. Thus the proportion of glycolysis and oxidative phosphorylation is maintained the same regardless of circadian time, and the pH difference between the tissue and superfusion solution can therefore be used to evaluate total energy production. We conclude that circadian clock regulation of retinal pH reflects circadian regulation of retinal energy metabolism.

scholarly journals Targeting Energy Metabolism in Mycobacterium tuberculosis, a New Paradigm in Antimycobacterial Drug Discovery

mBio ◽  
2017 ◽  
Vol 8 (2) ◽  
Author(s):  
Dirk Bald ◽  
Cristina Villellas ◽  
Ping Lu ◽  
Anil Koul
Keyword(s):  
Drug Discovery ◽  
Energy Metabolism ◽  
Oxidative Phosphorylation ◽  
Multidrug Resistant ◽  
New Paradigm ◽  
Mycobacterial Infections ◽  
Highly Active ◽  
Oxidative Phosphorylation Pathway ◽  
Novel Target ◽  
Synthase Inhibitor

ABSTRACT Drug-resistant mycobacterial infections are a serious global health challenge, leading to high mortality and socioeconomic burdens in developing countries worldwide. New innovative approaches, from identification of new targets to discovery of novel chemical scaffolds, are urgently needed. Recently, energy metabolism in mycobacteria, in particular the oxidative phosphorylation pathway, has emerged as an object of intense microbiological investigation and as a novel target pathway in drug discovery. New classes of antibacterials interfering with elements of the oxidative phosphorylation pathway are highly active in combating dormant or latent mycobacterial infections, with a promise of shortening tuberculosis chemotherapy. The regulatory approval of the ATP synthase inhibitor bedaquiline and the discovery of Q203, a candidate drug targeting the cytochrome bc 1 complex, have highlighted the central importance of this new target pathway. In this review, we discuss key features and potential applications of inhibiting energy metabolism in our quest for discovering potent novel and sterilizing drug combinations for combating tuberculosis. We believe that the combination of drugs targeting elements of the oxidative phosphorylation pathway can lead to a completely new regimen for drug-susceptible and multidrug-resistant tuberculosis.

scholarly journals Inhibition of glycerol metabolism in hepatocytes isolated from endotoxic rats

1997 ◽  
Vol 325 (2) ◽  
pp. 519-525 ◽  
Author(s):  
Pascale LECLERCQ ◽  
Céline FILIPPI ◽  
Brigitte SIBILLE ◽  
Sarah HAMANT ◽  
Christiane KERIEL ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Oxidative Phosphorylation ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Kinase Activity ◽  
Glycerol Metabolism ◽  
Transcription Rate ◽  
Intact Cells ◽  
Cellular Volume ◽  
Main Effect ◽  
Oxidative Phosphorylation Pathway

Sepsis or endotoxaemia inhibits gluconeogenesis from various substrates, the main effect being related to a change in the phosphoenolpyruvate carboxykinase transcription rate. In addition, sepsis has been reported to affect the oxidative phosphorylation pathway. We have studied glycerol metabolism in hepatocytes isolated from rats fasted and injected 16 h previously with lipopolysaccharide from Escherichiacoli. Endotoxin inhibited glycerol metabolism and led to a very large accumulation of glycerol 3-phosphate; the cytosolic reducing state was increased. Furthermore glycerol kinase activity was increased by 33% (P < 0.01). The respiratory rate of intact cells was significantly decreased by sepsis, with glycerol or octanoate as exogenous substrates, whereas oxidative phosphorylation (ATP-to-O ratio or respirations in state 4, state 3 and the oligomycin-insensitive state as well as the uncoupled state) was unchanged in permeabilized hepatocytes. Hence the effect on energy metabolism seems to be present only in intact hepatocytes. An additional important feature was the observation of a significant increase in cellular volume in cells from endotoxic animals, which might account for the alterations induced by sepsis.

scholarly journals Uridine Prevents Negative Effects of OXPHOS Xenobiotics on Dopaminergic Neuronal Differentiation

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1407
Author(s):  
Eldris Iglesias ◽  
M. Pilar Bayona-Bafaluy ◽  
Alba Pesini ◽  
Nuria Garrido-Pérez ◽  
Patricia Meade ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Oxidative Phosphorylation ◽  
Neuronal Differentiation ◽  
Fetal Brain ◽  
Global Dna Methylation ◽  
Protective Agent ◽  
Negative Consequences ◽  
Negative Effects ◽  
Negative Effect

Neuronal differentiation appears to be dependent on oxidative phosphorylation capacity. Several drugs inhibit oxidative phosphorylation and might be detrimental for neuronal differentiation. Some pregnant women take these medications during their first weeks of gestation when fetal nervous system is being developed. These treatments might have later negative consequences on the offspring’s health. To analyze a potential negative effect of three widely used medications, we studied in vitro dopaminergic neuronal differentiation of cells exposed to pharmacologic concentrations of azidothymidine for acquired immune deficiency syndrome; linezolid for multidrug-resistant tuberculosis; and atovaquone for malaria. We also analyzed the dopaminergic neuronal differentiation in brains of fetuses from pregnant mice exposed to linezolid. The drugs reduced the in vitro oxidative phosphorylation capacity and dopaminergic neuronal differentiation. This differentiation process does not appear to be affected in the prenatally exposed fetus brain. Nevertheless, the global DNA methylation in fetal brain was significantly altered, perhaps linking an early exposure to a negative effect in older life. Uridine was able to prevent the negative effects on in vitro dopaminergic neuronal differentiation and on in vivo global DNA methylation. Uridine could be used as a protective agent against oxidative phosphorylation-inhibiting pharmaceuticals provided during pregnancy when dopaminergic neuronal differentiation is taking place.

In vitro methylation of bovine papillomavirus alters its ability to transform mouse cells

1986 ◽  
Vol 6 (8) ◽  
pp. 2910-2915
Author(s):  
B A Christy ◽  
G A Scangos
Keyword(s):  
Escherichia Coli ◽  
Bovine Papillomavirus ◽  
Negative Effects ◽  
Absolute Level ◽  
Mouse Cells ◽  
The Absolute ◽  
In Cells

Bovine papillomavirus (BPV) was methylated in vitro at either the 29 HpaII sites, the 27 HhaI sites, or both. Methylation of the HpaII sites reduced transformation by the virus two- to sixfold, while methylation at HhaI sites increased transformation two- to fourfold. DNA methylated at both HpaII and HhaI sites did not differ detectably from unmethylated DNA in its efficiency of transformation. These results indicate that specific methylation sites, rather than the absolute level of methylated cytosine residues, are important in determining the effects on transformation and that the negative effects of methylation at some sites can be compensated for by methylation at other sites. BPV molecules in cells transformed by methylated BPV DNA contained little or no methylation, indicating that the pattern of methylation was not faithfully retained in these extrachromosomally replicating molecules. Methylation at the HpaII sites (but not the HhaI sites) in the cloned BPV plasmid or in pBR322 also inhibited transformation of the plasmids into Escherichia coli HB101 cells.

Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

1994 ◽  
Vol 52 ◽  
pp. 270-271
Author(s):  
Henry H. Eichelberger ◽  
John G. Baust ◽  
Robert G. Van Buskirk
Keyword(s):  
Cold Storage ◽  
Structural Integrity ◽  
Culture Media ◽  
Cell Structure ◽  
Natural State ◽  
Sodium Cacodylate ◽  
Ruthenium Tetroxide ◽  
Cacodylate Buffer ◽  
Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

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