Dose-dependence of antiapoptotic and toxic action of ouabain in neurons of primary cultures of rat cortex

Bolshakov, A. E.; Сибаров, Д. А.; Абушик, П. А.; Кривой, И. И.; Антонов, С. М.

doi:10.1134/s1990747812050029

Cited by 7 publications

(10 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The excitotoxic stress response starts with a free intracellular Ca 2+ overload which is the most important of apoptosis (Khodorov, 2004 ). A similar mechanism of neuronal dysfunction and cell apoptosis is induced by micromolar ouabain (Kulikov et al, 2007 ; Bolshakov et al, 2012 ). However, some animal experimental studies have shown that low-doses of CTS provide neuroprotection against ischemia (Wang et al, 2006 ; Oselkin et al, 2010 ).…”

Section: Cardiotonic Steroids and Cell Survivalmentioning

confidence: 90%

“…Recently it was shown that ouabain at subnanomolar concentrations can prevent GluR agonist-induced apoptosis in primary culture of rat cortical neurons (Bolshakov et al, 2012 ; Sibarov et al, 2012 ). Apoptotic injury was prevented when the agonists were applied together with 0.1–1 nM ouabain resulting in survival of neurons in this model of excitotoxicity.…”

Section: Cardiotonic Steroids and Cell Survivalmentioning

confidence: 99%

“…In addition, the postsynaptic epileptiform currents, reflecting periodical asynchronous glutamate release associated with elevations in intracellular Ca 2+ concentration, were found to be suppressed by 1 nM ouabain (Sibarov et al, 2014 ). Ouabain was, however, found to have a bimodal effect; including anti-apoptotic action in excitotoxic stress in the concentration range from 0.1 to 1 nM, and toxic action at concentrations 10 nM–30 μM (Bolshakov et al, 2012 ).…”

Section: Cardiotonic Steroids and Cell Survivalmentioning

confidence: 99%

See 2 more Smart Citations

Specialized Functional Diversity and Interactions of the Na,K-ATPase

Matchkov

Кривой

2016

Front. Physiol.

View full text Add to dashboard Cite

Na,K-ATPase is a protein ubiquitously expressed in the plasma membrane of all animal cells and vitally essential for their functions. A specialized functional diversity of the Na,K-ATPase isozymes is provided by molecular heterogeneity, distinct subcellular localizations, and functional interactions with molecular environment. Studies over the last decades clearly demonstrated complex and isoform-specific reciprocal functional interactions between the Na,K-ATPase and neighboring proteins and lipids. These interactions are enabled by a spatially restricted ion homeostasis, direct protein-protein/lipid interactions, and protein kinase signaling pathways. In addition to its “classical” function in ion translocation, the Na,K-ATPase is now considered as one of the most important signaling molecules in neuronal, epithelial, skeletal, cardiac and vascular tissues. Accordingly, the Na,K-ATPase forms specialized sub-cellular multimolecular microdomains which act as receptors to circulating endogenous cardiotonic steroids (CTS) triggering a number of signaling pathways. Changes in these endogenous cardiotonic steroid levels and initiated signaling responses have significant adaptive values for tissues and whole organisms under numerous physiological and pathophysiological conditions. This review discusses recent progress in the studies of functional interactions between the Na,K-ATPase and molecular microenvironment, the Na,K-ATPase-dependent signaling pathways and their significance for diversity of cell function.

show abstract

Section: Cardiotonic Steroids and Cell Survivalmentioning

confidence: 90%

Section: Cardiotonic Steroids and Cell Survivalmentioning

confidence: 99%

Section: Cardiotonic Steroids and Cell Survivalmentioning

confidence: 99%

See 1 more Smart Citation

Specialized Functional Diversity and Interactions of the Na,K-ATPase

Matchkov

Кривой

2016

Front. Physiol.

View full text Add to dashboard Cite

show abstract

“…Agonists were also combined with 0.1 or 1 nM oubain. These concentrations of ouabain provided neuroprotection against exciotoxicity when studied for dose-dependence [31]. Here both glutamate receptor agonists were co-applied with 30 µM glycine to ensure activation of NMDA receptors.…”

Section: Quantitation Of Cell Viabilitymentioning

confidence: 99%

Calcium Export from Neurons and Multi-Kinase Signaling Cascades Contribute to Ouabain Neuroprotection in Hyperhomocysteinemia

et al. 2020

View full text Add to dashboard Cite

Pathological homocysteine (HCY) accumulation in the human plasma, known as hyperhomocysteinemia, exacerbates neurodegenerative diseases because, in the brain, this amino acid acts as a persistent N-methyl-d-aspartate receptor agonist. We studied the effects of 0.1–1 nM ouabain on intracellular Ca2+ signaling, mitochondrial inner membrane voltage (φmit), and cell viability in primary cultures of rat cortical neurons in glutamate and HCY neurotoxic insults. In addition, apoptosis-related protein expression and the involvement of some kinases in ouabain-mediated effects were evaluated. In short insults, HCY was less potent than glutamate as a neurotoxic agent and induced a 20% loss of φmit, whereas glutamate caused a 70% decrease of this value. Subnanomolar ouabain exhibited immediate and postponed neuroprotective effects on neurons. (1) Ouabain rapidly reduced the Ca2+ overload of neurons and loss of φmit evoked by glutamate and HCY that rescued neurons in short insults. (2) In prolonged 24 h excitotoxic insults, ouabain prevented neuronal apoptosis, triggering proteinkinase A and proteinkinase C dependent intracellular neuroprotective cascades for HCY, but not for glutamate. We, therefore, demonstrated here the role of PKC and PKA involving pathways in neuronal survival caused by ouabain in hyperhomocysteinemia, which suggests existence of different appropriate pharmacological treatment for hyperhomocysteinemia and glutamate excitotoxicity.

show abstract

“…Neurons express α1and α3-isoforms of NKA. NKAα1 is many-folds less sensitive to ouabain than NKAα3, which is inhibited by ouabain at concentrations above 10 nM [30][31]. Ouabain at 1 nM, however, potentiates ion transport by NKAα3 without any effect on NKAα1 [30].…”

Section: Introductionmentioning

confidence: 96%

Calcium export from neurons and multi-kinase signaling cascades contribute to ouabain neuroprotection in hyperhomocysteinemia

Ivanova

Kokorina

Timofeeva

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Subnanomolar ouabain binding to the Na,K-ATPase triggers intracellular signaling, prevents an overload of neurons with Ca2+, and their apoptosis caused by glutamate receptor agonists. Elevated plasma homocysteine (HCY), known as hyperhomocysteinemia, represents a risk factor for stroke and can exacerbate many neuronal disorders. HCY acts as a persistent N-methyl-D-aspartate receptor (NMDAR) agonist, which, in contrast to glutamate, desensitizes NMDARs containing GluN2B subunits. Mechanisms of HCY neurotoxicity remain not clearly understood since GluN2B-containing NMDARs provide a major contribution to excitotoxicity among glutamate receptors.Methods: Using fluorescent tools combined with the confocal microscopy, we compared 0.1 - 1 nM ouabain effects on the intracellular Ca2+ signaling, on the mitochondrial inner membrane voltage and the cell viability in primary cultures of rat cortical neurons in glutamate and HCY neurotoxic insults. We also studied an apoptosis-related protein expression and the involvement of some kinases in ouabain mediated effects.Results: In short insults HCY was less potent than glutamate as a neurotoxic agent. This amino acid induced the voltage loss (∆φmit) of 0.2 of the total mitochondrial inner membrane voltage (φmit) instead of ∆φmit = 0.7 for glutamate. We have found that subnanomolar ouabain exhibited rapid and postponed neuroprotective effects on neurons and (1) rapidly reduced the Ca2+ overload of neurons and the voltage loss of inner mitochondrial membranes evoked by glutamate and HCY, and (2) prevented neuronal apoptosis during 24 h treatments with glutamate or HCY. Using a set of specific kinase inhibitors such as PKA inhibitor, chelerythrine, and KN93, we demonstrated the role of multi-kinase signaling pathways involving PKC and PKA in neuronal survival caused by ouabain in hyperhomocysteinemia. Conclusions: Subnanomolar ouabain prevents neurodegeneration caused by glutamate and HCY. For both amino acids, ouabain evokes an acceleration of Ca2+ export by sodium-calcium exchangers from neurons preventing the voltage loss by mitochondrial inner membranes that rescue neurons in short insults. In prolonged insults, ouabain triggers intracellular neuroprotective cascades, including activation of PKA and PKC for HCY, but not for glutamate. This suggests that different appropriate pharmacology for hyperhomocysteinemia and glutamate excitotoxicity could be applied for clinical treatments.

show abstract

Dose-dependence of antiapoptotic and toxic action of ouabain in neurons of primary cultures of rat cortex

Cited by 7 publications

References 20 publications

Specialized Functional Diversity and Interactions of the Na,K-ATPase

Specialized Functional Diversity and Interactions of the Na,K-ATPase

Calcium Export from Neurons and Multi-Kinase Signaling Cascades Contribute to Ouabain Neuroprotection in Hyperhomocysteinemia

Calcium export from neurons and multi-kinase signaling cascades contribute to ouabain neuroprotection in hyperhomocysteinemia

Contact Info

Product

Resources

About