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N K Isaev

Publications and source records attributed to N K Isaev.

16 recordsLinked to original sources
PubMed2000

Inhibition of Na(+),K(+)-ATPase activity in cultured rat cerebellar granule cells prevents the onset of apoptosis induced by low potassium.

In cerebellar granule cells in culture, lowering of extracellular [K(+)] results in apoptotic death (D'Mello, S.R., Galli, C., Ciotti, T. and Calissano, P., Induction of apoptosis in cerebellar granule neurons by low potassium: inhibition of death by insulin-like growth factor I and cAMP, Proc. Natl. Acad. Sci. USA, 90 (1993) 10989-10993). In this model, we studied the influence of Na(+), K(+)-ATPase inhibition on apoptosis. We demonstrate that cell death (93+/-2 vs. 46+/-1.6%) as well as fragmentation of nuclear DNA induced by low extracellular potassium were prevented by addition of ouabain (0.1 mM), a specific inhibitor of the Na(+),K(+)-ATPase. Blockade of glutamatergic N-methyl-D-aspartate and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors by 5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclohepten-5,10-imine hydrogen maleate (MK-801; 20 microM) and 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX; 50 microM) did not inhibit the protective effect of ouabain. 24 h treatment with ouabain also decreased cell death induced by Fe(2+)/ascorbic acid (74+/-2% to 49+/-3%). We speculate that ouabain pretreatment enhances the resistance against low [K(+)]-induced apoptosis independent of glutamate-receptor activation. Since this effect can be mimicked by a free-radical generating system, we suggest an antioxidative effect underlying ouabain-induced neuroprotection.

Adenosine Triphosphatases↗
PubMed1999

Induction of ischemic tolerance in rat cortical neurons by 3-nitropropionic acid: chemical preconditioning.

Sublethal ischemia leads to increased tolerance against subsequent ischemia. We investigated whether tolerance could also be elicited by mild respiratory-chain inhibition (chemical hypoxia) in a rat neuronal-cell enriched culture system. 3-Nitropropionic acid (3-NPA) caused a concentration-dependent inhibition of succinate-dehydrogenase. Two hours preconditioning with 3-NPA 24-48 h before oxygen-glucose deprivation (OGD) reduced neuronal damage morphologically and reduced lactate deydrogenase (LDH) release up to 72% compared to sham-treated sister cultures without 3-NPA. In an attempt to elucidate transcriptional mechanisms, we found no rapid translocation of the hypoxia-sensitive transcription factors N F-KB or hypoxia-inducible factor-I (HIF-I) at 3-NPA concentrations sufficient to trigger tolerance against OGD. In accordance to previous in vivo and brain slice data, we conclude that 3-NPA chemically induces tolerance against oxygen-glucose deprivation in vitro. However, the underlying mechanisms remain elusive.

Animals↗
PubMed1998

The lack of extracellular Na+ exacerbates Ca2+-dependent damage of cultured cerebellar granule cells.

Rhodamine 123 staining, light and electron microscopy were used to evaluate the ultrastructural and functional state of cultured cerebellar granule cells after short treatment with the solution where NaCl was substituted by sucrose (sucrose balance salt medium, SBSM). Cell exposure to SBSM for 20 min resulted in the fact that mitochondria in the neurons lost their ability to sequester rhodamine 123. This effect could be prevented by: (i) non-competitive N-methyl-D-aspartate (NMDA) receptor channel blocker, 10(-5) M MK-801; (ii) a competitive specific antagonist of NMDA glutamate receptors, 0.25 x 10(-3) M D,L-2-amino-7-phosphonoheptanoate (APH); (iii) 10(-3) M cobalt chloride; (iv) removal of Ca2+ from the medium. Low Na+ in the Ca2+-containing medium caused considerable mitochondrial swelling in granule cells. However, the same treatment in the absence of calcium ions in the medium abolished the deleterious effect of SBSM on the neuronal mitochondrial structure and functions. It is suggested that (i) the exposure of cultured cerebellar granule cells to SBSM leads to a release of endogenous glutamate from cells; (ii) Ca2+ ions potentially de-energizing neuronal mitochondria enter the neuron preferentially through the NMDA channels rather than through the Na+/Ca2+ exchanger; (iii) mitochondrial swelling in granule cells is highly Ca2+-dependent; (iv) cellular overload with sodium ions can activate mitochondrial Na+/Ca2+ exchanger and thus prevent permeability transition pore opening in mitochondria.

Animals↗
PubMed1997

Induction of tolerance in rat cortical neurons: hypoxic preconditioning.

Sublethal ischemia leads to increased tolerance against subsequent prolonged cerebral ischemia in vivo. In the present study we modeled preconditioning mechanisms in a neuronal-enriched culture. Damage was significantly reduced (up to 72%) with 1.5 h of oxygen-glucose deprivation 48-72 h before 3 h oxygen-glucose deprivation. Tolerance was also elicited by Na+-K+-ATPase inhibition. No damage was observed when astroglial or endothelial cells were exposed to hypoxia for 3 and 6 h, respectively. We conclude that hypoxic preconditioning is a robust neuronal phenomenon in vitro with a similar temporal pattern and selective cellular vulnerability as the ischemic tolerance phenomenon shown in vivo.

Animals↗
PubMed1996

Neurotoxic glutamate treatment of cultured cerebellar granule cells induces Ca2+ -dependent collapse of mitochondrial membrane potential and ultrastructural alterations of mitochondria.

Rhodamine 123 staining and electron microscopy were used to reveal a correlation between the ultrastructural and functional state of cultured cerebellar granule cells after short glutamate treatment. Glutamate exposure (15 min, 100 microM) in Mg2+-free solution caused considerable ultrastructural alterations in a granule cell: clumping of the chromatin, swelling of the endoplasmic reticulum and mitochondria, and disruption of the mitochondrial cristae. After glutamate treatment, the mitochondria of the neurons lost their ability to sequester rhodamine 123. Both the N-methyl-D-aspartate receptor channel blocker MK-801 (30 microM) and cobalt chloride (2 mM) prevented the deteriorative effects of glutamate. These data suggest that glutamate-induced Ca2+ overload of the neurons can lead to non-specific permeability of the inner mitochondrial membrane, resulting in neuronal death.

Animals↗
PubMed1992

[Effects of nerve growth factor on the development of the dendritic system of cholinergic neurons in dissociated culture of the rat septum].

Neurons dissociated from septal area of fetal (E18-19) rat brain were grown 14-days in culture. Cholinergic neurons were identified by cytochemical demonstration of acetyl cholinesterase. It was shown that the nerve growth factor added to the culture medium (50 u/ml) has increased the size of cell body of AchE-positive neurons, mean total length and arborization of dendrites and also the dendritic tree area.

Animals↗
PubMed1991

[The effect of nerve growth factor on cholinergic neurons in dissociated cultures of the septum pellucidum].

Neurons dissociated from septal area of fetal (E 18-19) rat brain were grown for 7 days in culture. Cholinergic neurons were identified by cytochemical demonstration of acetyl cholinesterase. Addition of the nerve growth factor to the culture medium (200 u/ml) increases the number of AChE-positive neurons, the size of the cell body and activity of AChE in these neurons.

Acetylcholinesterase↗
PubMed1990

[The use of a polyethyleneimine substrate for culturing dissociated cells of the central nervous system].

A new method of preparation of polyethylenimin substrate for cultivation of dissociated cells from different parts of the central nervous system is presented. The alcohol solution of polyethylenimin used for this substrate is easily prepared, needs no sterilization and can be stored unlimitedly. The substrate provides a good attachment, growth and differentiation of neurons and glial cells and possesses all the advantages of more expensive polyaminoacid substrates (polylysine, polyornithine). Our substrate is highly adhesive, displays good optical properties and no artefacts after histological and histochemical treatment.

Animals↗