Delayed Neuronal Death After Brief Histotoxic Hypoxia In Vitro

Uto, Akira; Dux, E.; Kusumoto, M.; Hossmann, K.‐A.

doi:10.1046/j.1471-4159.1995.64052185.x

Cited by 36 publications

(22 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Transient cytotoxic hypoxia allows recovery to a certain degree; however, independent from the NaCN concentration used, a decrease in viability was measured within the first 6 h. Nevertheless there is also clear evidence of regeneration when NaCN concentrations in a range from 0.01 mM to 1 mM are used. In contrast to other findings describing only partial neuronal degeneration of rat neuronal cultures due to a 10 mM cyanide lesion (Uto et al 1995), this dosage leads to an irreversible damage of chick cortical neurons without any signs of recovery.…”

Section: Discussioncontrasting

confidence: 97%

Effects of NaCN and ionomycin on neuronal viability and on the abundance of microtubule-associated proteins MAP1, MAP2, and tau in isolated chick cortical neurons

Hutter‐Paier¹,

Grygar²,

Loibner³

et al. 2000

Cell and Tissue Research

View full text Add to dashboard Cite

The effects of two toxins, sodium cyanide (NaCN) and ionomycin (IM), on neuronal viability and on the expression of the microtubule-associated proteins MAP1, MAP2, and tau were studied in isolated chick cortical neurons. Cytotoxic hypoxia due to NaCN treatment was performed to mimic acute neuronal damage, whereas long-term IM treatment was used as a model for chronic neuronal impairment. After 5 days in vitro, a cytotoxic lesion was induced either by addition of NaCN (0.01-10 mM) or IM (0.01-10 microM). The NaCN solution was aspirated after 30 min and cells were allowed to regenerate for 6 h, 24 h, 48 h, or 72 h; whereas the permanent IM lesions were left undisturbed during the same periods of time. Neuronal viability was assessed by MTT assay. The abundance of MAP1, MAP2, and tau was evaluated by immunoblotting and, for MAP2, by immunohistochemistry also. Results showed that NaCN and IM lesions dose-dependently decreased viability. Irreversible cell damage occurred after impairment with 10 mM NaCN and 1 microm or 10 microm IM, while neurons lesioned with lower concentrations regenerated partially or adapted to the toxic environment. However, the same level of viability as of untreated cells was never reached. Furthermore abundance of MAPs was changed after both lesions. But while after extended recovery from NaCN lesion protein expression was normalizing (MAP2) or at least still detectable (MAP1A, tau), the consequences of a permanent IM lesion were more severe, since neurons were not able to maintain or even restore their MAP expression. Immunohistochemical experiments for MAP2 revealed that, compared with controls, NaCN and, to a much higher extent, IM treatment resulted in a loss of immunoreactivity in neurites due to progressing cell death.

show abstract

Section: Discussioncontrasting

confidence: 97%

Effects of NaCN and ionomycin on neuronal viability and on the abundance of microtubule-associated proteins MAP1, MAP2, and tau in isolated chick cortical neurons

Hutter‐Paier¹,

Grygar²,

Loibner³

et al. 2000

Cell and Tissue Research

View full text Add to dashboard Cite

show abstract

“…Early studies showed that 30-min treatment of PC12 cells with cyanide reduced intracellular ATP concentrations ([ATP] i ) by 92% [Carroll et al, 1992]. Iodoacetate inhibited [ATP] i by 40% in cortical neuron cultures [Uto et al, 1995]. Sodium azide combined with acidosis caused a near-complete cell ATP depletion in astrocytes [Swanson et al, 1997].…”

Section: Discussionmentioning

confidence: 99%

“…Additional effects by iodoacetate and cyanide have been reported. Uto et al [1995] demonstrated that the delayed neuronal death induced by iodoacetate and cyanide was probably mediated by free-radicals. Furthermore, iodoacetate, but not cyanide, produced marked increase in plasma membrane permeability to Na ϩ , K ϩ , Pi and small molecules such as lucifer yellow [Kempson et al, 1991], and the release of preincorporated […”

Section: Discussionmentioning

confidence: 99%

ATP-stimulated c-fos andzif268 mRNA expression is inhibited by chemical hypoxia in a rat brain-derived type 2 astrocyte cell line, RBA-2

Hung¹,

Huang

Tsay³

et al. 2000

J. Cell. Biochem.

View full text Add to dashboard Cite

The stimulus-transcriptional coupling during ischemia/hypoxia was examined for ATP-stimulated expression of immediate early genes (IEGs; c-fos, zif268, c-myc and nur77) in a rat brain-derived type 2 astrocyte cell line, RBA-2. Incubation of cells with 1 mM of extracellular ATP stimulated time-dependent expression of c-fos and zif268. ATP induced the largest increases in zif268 mRNA and a lesser one in c-fos mRNA. ATP also induced a slight increase in nur77 mRNA but was ineffective in inducing c-myc expression in these cells. Brief exposure of cells to potassium cyanide to simulate chemical hypoxia induced 9-fold and 7-fold transient increases in c-fos and zif268 expression, respectively, but did not affect c-myc or nur77 expression. When cyanide and ATP were added together, the expression of c-fos and zif268 expression was inhibited, and the effect was mimicked by simulating chemical hypoxia with sodium azide. To elucidate the mechanism involved, the effect of cyanide on ATP-stimulated increases in intracellular Ca(2+) concentrations, [Ca(2+)](i), and phospholipase D (PLD) activities were measured. Cyanide induced an increase in [Ca(2&plus);](i) and further enhanced the ATP-stimulated increases in [Ca(2+)](i) and PLD activities. Nevertheless, metabolic inhibitor, iodoacetate, blocked the ATP-induced c-fos and partially inhibited zif268 expression, and deprivation of cells with glucose also inhibited the ATP-induced c-fos expression. Taken together, these results demonstrate that both extracellular ATP and chemical hypoxia induce c-fos and zif268 expression in RBA-2 type 2 astrocytes. The chemical hypoxia inhibited ATP-stimulated c-fos and zif268 expression is not due to alterations in Ca(2+) and PLD signaling, and is at least partially related to metabolic disturbance in these cells.

show abstract

“…In other respects, several lines of evidence arising from both in vivo and in vitro studies suggest the participation of an apoptotic cascade in delayed brain injury observed after hypoxia-ischemia (Beilharz et al, 1995;Charriaut-Marlangue et al, 1996), whereas excitotoxicity resulting from exposure to excess glutamate has been associated with both cell necrosis (Dessi et al, 1993;Chihab et al, 1998b) and apoptosis (Bonfoco et al, 1995;Wood and Bristow, 1998). However, very few studies have been devoted to the consequences of hypoxia alone, as the implication of glutamate is more controversial (Aitken et al, 1988;Uto et al, 1995;Chihab et al, 1998a;Chow and Haddad, 1998;Banasiak et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

“…whereas others found major differences between the effects of glutamate and those of hypoxia or anoxia (Aitken et al, 1988;Uto et al, 1995;Chow and Haddad, 1998a). In our study, administration of the NMDA receptor antagonist, MK-801, to newborn pups before exposure to hypoxia did not reduce neuronal injury in the CA1 hippocampus.…”

mentioning

confidence: 99%

Differential neuronal fates in the CA1 hippocampus after hypoxia in newborn and 7‐day‐old rats: Effects of pre‐treatment with MK‐801

et al. 2003

View full text Add to dashboard Cite

The brain displays an age-dependent sensitivity to ischemic insults. However, the consequences of oxygen deprivation per se in the developing brain remain unclear, and the role of glutamate excitotoxicity via N-methyl-D-aspartate (NMDA) receptors is controversial. To gain a better understanding of the mechanisms involved in the cerebral response to severe hypoxia, cell damage was temporally monitored in the CA1 hippocampus of rat pups transiently exposed to in vivo hypoxia (100% N2) at either 24 h or 7 days of age. Also, the influence of a pre-treatment with the NMDA receptor antagonist MK-801 (5 mg/kg, i.p.) was examined. At both ages, morphometric analyses and cell counts showed hypoxia-induced significant neuronal loss (30-35%) in the pyramidal layer, with injury appearing more rapidly in rats exposed at 7 days. Morphological alterations of 4,6-diamidino-2-phenylindole (DAPI)-labeled nuclei, DNA fragmentation patterns on agarose gels, as well as expression profiles of the apoptosis-related regulatory proteins Bax and Bcl-2 showed that apoptosis was prevalent in younger animals, whereas only necrosis was detected in hippocampi of rats treated at 7 days. Moreover, pre-treatment with MK-801 was ineffective in protecting hippocampal neurons from hypoxic injury in newborn rats, but significantly reduced necrosis in older subjects. These data confirm that hypoxia alone may trigger neuronal death in vivo, and the type of cell death is strongly influenced by the degree of brain maturity. Finally, NMDA receptors are not involved in the apoptotic consequences of hypoxia in the newborn rat brain, but they were found to mediate necrosis at 7 days of age.

show abstract

Delayed Neuronal Death After Brief Histotoxic Hypoxia In Vitro

Cited by 36 publications

References 0 publications

Effects of NaCN and ionomycin on neuronal viability and on the abundance of microtubule-associated proteins MAP1, MAP2, and tau in isolated chick cortical neurons

Effects of NaCN and ionomycin on neuronal viability and on the abundance of microtubule-associated proteins MAP1, MAP2, and tau in isolated chick cortical neurons

ATP-stimulated c-fos andzif268 mRNA expression is inhibited by chemical hypoxia in a rat brain-derived type 2 astrocyte cell line, RBA-2

Differential neuronal fates in the CA1 hippocampus after hypoxia in newborn and 7‐day‐old rats: Effects of pre‐treatment with MK‐801

Contact Info

Product

Resources

About