Biomedical subjects

J Krieglstein

Publications and source records attributed to J Krieglstein.

At least 91 records · Page 5Linked to original sources

PubMed1995

Quantitative analysis of the electrocorticogram after forebrain ischemia in the rat.

The purpose of the study was to test the hypothesis that the postischemic neuronal damage is accompanied by changes of the electrocorticogram (ECoG) of freely moving rats during long-term recovery after forebrain ischemia. Ten minutes forebrain ischemia was induced in male Wistar rats. ECoG was recorded 1 day before as well as 1 h, 1 day and 7 days after ischemia. The ECoG power was calculated and for characterizing postischemic ECoG development the percentage of preischemic ECoG power was evaluated. The awake state of the rats was considered for analysis only. Furthermore, the neuroprotective effect of phencyclidine (PCP, 5 mg/kg i.v., 15 min prior to ischemia) was demonstrated in the ECoG changes. One hour after ischemia the ECoG power of the theta (4.75-6.75 Hz), alpha (7.00-12.50 Hz) and beta (12.75-18.50 Hz) band was decreased compared with sham-operated controls and PCP did not influence these changes. However, 1 day after ischemia ECoG power of the saline-treated ischemic rats was completely restored and no longer different from that of the sham-operated group. PCP-treated ischemic animals showed significantly elevated ECoG power in comparison with saline-treated animals. Seven days after ischemia ECoG power of the saline-treated ischemic rats again decreased. In comparison with the ischemic controls, the ECoG activity of the PCP-treated ischemic rats was significantly higher. PCP maintained postischemic ECoG power at the non-ischemic control level. It is suggested that the decrease in the ECoG activity several days after ischemia is related to the delayed neuronal damage.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

Strain differences in vulnerability of hippocampal neurons to transient cerebral ischaemia in the rat.

Strain differences in the vulnerability of hippocampal neurons to an ischaemic insult were investigated in Sprague-Dawley, Wistar and Fischer 344 rats. Transient global brain ischaemia was produced for 5 minutes by a combination of bilateral carotid artery occlusion and oligaemic hypotension (40 mmHg) induced by exsanguination. The number of viable neurons in the CA1 subfield was counted under a light microscope 7 days after the ischaemic insult. The density of viable neurons in the CA1 subfield of normal rats was around 150 cells/mm of CA1 length in all the strains examined; after global brain ischaemia, this parameter in Sprague-Dawley, Wistar and Fischer 344 strain rats was approximately 110, 120, and 70, respectively. The results suggest that the hippocampal CA1 neurons of Fischer 344 strain rats are more vulnerable to ischaemic insult than those of the other strains. There were many F344 rats (8/21) which showed atypical vasculature patterns in the posterior region of the circle of Willis, suggesting less blood flow through anastomosis from basilar artery to posterior cerebral artery and/or ill-balanced blood flow between left and right hemispheres. These anatomical variations in the circle of Willis may in part contribute to the strain differences in the vulnerability to cerebral ischaemia.

Animals↗

PubMed1994

Over-additive protective effect of dizocilpine and NBQX against neuronal damage.

Several reports have indicated that the two glutamate receptor antagonists, dizocilpine (that binds to the phencyclidine recognition site of the NMDA (N-methyl-D-aspartate) receptor) and NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline, that binds to the AMPA (alpha-amino-3-hydroxy-5-methyl-isoxazole) receptor), protect neurons against damage caused by hypoxia, ischemia or excitotoxicity. We, therefore, used a combination of these drugs to achieve enhanced neuroprotection. Primary cultures of rat hippocampal neurons were challenged by glutamate intoxication. Both dizocilpine and NBQX produced dose-dependent increases in the percentage of viable neurons. Combined treatment with both glutamate receptor antagonists had an over-additive neuroprotective effect. Simultaneous administration of dizocilpine and NBQX also had a pronounced neuroprotective effect in vivo in mice subjected to focal cerebral ischemia and rats with global forebrain ischemia. This suggest that such a combination may have therapeutic relevance.

Animals↗

PubMed1994

Primary cultures of neurons for testing neuroprotective drug effects.

Primary cultures of neurons are widely used for the investigation of pathomechanisms of neuronal damage and for the evaluation of neuroprotective drug effects. The present paper gives a short survey of frequently used primary neuronal culture systems and of experimental measures for the induction of defined neuronal damage with particular respect to the pathomechanisms of cerebral ischemia. Neuroprotective drug effects as achieved under these conditions are reviewed, and the neuroprotective effects of glutamate antagonists, radical scavengers, and neural growth factors are discussed in some more detail.

Animals↗

PubMed1994

Protective effects of 5-HT1A receptor agonists against neuronal damage demonstrated in vivo and in vitro.

The aim of the present study was to evaluate the neuroprotective effect of the 5-hydroxytryptamine1A (5-HT1A) agonists, CM 57493 and urapidil, in vivo and in vitro, respectively. In vivo permanent occlusion of the middle cerebral artery (MCA) was performed in male Wistar rats. Forty-eight hours after electrocoagulation of the MCA the infarct volume was determined. Pretreatment of the rat with the 5-HT1A agonist urapidil significantly reduced infarct development. The neuroprotective effect of the agent was restricted to the cortical area; the striatal damage was not influenced. As the stimulation of the 5-HT1A receptor by serotonin is supposed to induce inhibitory, hyperpolarizing effects by opening of a Ca(2+)-independent neuronal K+ ionophore, the efficacy of agonistic drugs directly on the neuron was investigated in vitro. Cyanide-induced cytotoxic hypoxia as well as glutamate-induced excitotoxicity were performed using primary neuronal cell cultures from chick embryo cerebral hemispheres. Treatment with the 5-HT1A agonists urapidil and CM 57493 significantly increased protein content of hypoxic cultures. CM 57493 added to the culture medium (1-10 microM) during and up to 24 h after glutamate exposure ameliorated viability of the neurons. The results demonstrate neuroprotective potency of the 5-HT1A agonists, urapidil and CM 57493, when applied under hypoxic, excitotoxic and ischemic conditions in vivo and in vitro, respectively. Both, presynaptically induced inhibition of glutamate release as well as postsynaptically induced inhibition of neuronal excitability could be discussed as possible mechanisms of action of the 5-HT1A receptor agonism.

Animals↗

PubMed1994

Neuronal protective effects of calcium antagonists in cerebral ischemia.

We investigated the effects of calcium antagonists against ischemic injury in vivo and against excitotoxic damage in vitro. In vivo nimodipine protected significantly the CA1 hippocampal neurons from neurodegeneration after transient global ischemia in rats without changing the local cerebral blood flow. Furnidipine reduced the area of ischemia after permanent MCA-occlusion in mice. The results in vitro using the Ca(2+)-sensitive dye fura-2 showed that nimodipine reduced in a dose-dependent manner the elevation of [Ca2+]i in hippocampal neurons induced by K(+)-stimulation. The present in vitro and in vivo data show that calcium antagonists are potent agents in protecting neurons against the deleterious consequences of an excitotoxic or ischemic insult.

Animals↗

PubMed1994

Opposing effects of transforming growth factor-beta 1 on glutamate neurotoxicity.

Activation of microglia has been emphasized as a critical step in the pathophysiology of degenerative and inflammatory processes of the CNS. Activated microglia release low molecular weight compounds, such as excitatory amino acids, that are directly toxic to neurons. Here we demonstrate that a microglia-derived cytokine, transforming growth factor-beta 1, directly alters the susceptibility of neurons to glutamate-induced cell damage. Transforming growth factor-beta 1 acts as a neuroprotectant following short-term exposure to glutamate, whereas, following chronic exposure to glutamate, similar concentrations of transforming growth factor-beta 1 actually potentiate excitotoxic cell death. This complex interaction may play an important role in determining the extent of local tissue damage.

Animals↗

PubMed1994

Extension of the 2-deoxyglucose method to the fetus in utero: theory and normal values for the cerebral glucose consumption in fetal guinea pigs.

Fetal cerebral metabolism changes during development. The normal fetal metabolic rate must be known to evaluate pathophysiological changes. Therefore, we determined the regional cerebral glucose consumption in the fetal guinea pig. This required the application of the 2-deoxyglucose method to this species. We measured both the transfer coefficients of deoxyglucose and glucose between the maternal arterial plasma and the fetal brain and the lumped constant in chronically prepared undisturbed guinea pig dams using a three-compartment model. Furthermore, the ratio between the initial clearances of deoxyglucose and glucose between the maternal arterial plasma and the fetal brain and the ratio between the phosphorylation coefficients of these substrates in the fetal brain were determined. The total cerebral glucose consumption measured by the deoxyglucose method (10 +/- 1.2 mumol/100 g/min) was similar to that calculated from the glucose concentration and the phosphorylation coefficient of glucose in the cerebrum (10 +/- 0.4 mumol/100 g/min). We conclude that the 2-deoxyglucose method is applicable to the guinea pig, and we further conclude that in the fetal guinea pig cerebral glucose consumption is 10 times lower than that in the adult.

Animals↗

PubMed1994

Neuroprotective effects of TGF-beta 1.

The transforming growth factor-beta 1 (TGF-beta 1) has been shown to be an injury-related peptide growth factor within the mammalian brain. We tested TGF-beta 1 for its protective effects against neuronal degeneration caused by sodium cyanide (1 mM) or by the excitatory amino acid L-glutamate (1 mM) in vitro and against ischemic injury in vivo caused by permanent occlusion of the left middle cerebral artery in mice. In vitro, TGF-beta 1 (1-30 ng/ml) significantly reduced hypoxic and excitotoxic neuronal damage in a concentration-dependent manner. In vivo, intracerebroventricular administration of TGF-beta 1 (1 microgram/kg) decreased the infarcted area on the mouse brain surface. The present results suggest that TGF-beta 1 is capable of protecting neurons against damage both in vitro and in vivo.

Animals↗

PubMed1993

Effects of serotonergic drugs in experimental brain ischemia: evidence for a protective role of serotonin in cerebral ischemia.

We have examined the significance of the serotonergic system in the pathophysiology of ischemic brain damage. Permanent occlusion of the middle cerebral artery (MCA) was performed in male NMRI mice. After 48 h, the animals received a transcardiac injection of carbon black. The area of ischemia was restricted to the neocortex and its size was determined planimetrically by means of an image analyzing system. In control experiments, the NMDA antagonist dizocilpine (MK-801), the AMPA/kainate antagonist NBQX (2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(F)-quinoxaline) and the L-type calcium channel blocker nimodipine all produced a significant reduction in ischemic injury of the mouse neocortex. Interestingly, all of the 5-HT1A agonists tested (ipsapirone, CM 57493 [4-(3-trifluoromethylphenyl)-1-(2-cyanoethyl)-1,2,3,6-tetrahydropyridine ] and urapidil) were equally efficacious in reducing ischemic injury. On the other hand, the 5-HT2 antagonist naftidrofuryl failed to protect the brain tissue significantly against ischemic brain damage. Roxindole, a 5-HT1A agonist and 5-HT uptake inhibitor, was the most potent serotonergic compound tested. In order to examine the effects of 5-HT1A receptor activation in a different context, 10 min of forebrain ischemia was induced in male Wistar rats by a bilateral occlusion of the common carotid arteries combined with systemic hypotension. Administration of the 5-HT1A agonist CM 57493 reduced the neuronal damage within the ventral hippocampus and the entorhinal cortex as assessed histologically 7 days after ischemia. Finally, we found that 5-HT1A agonists are capable of reducing neuronal damage of cultured neocortical and hippocampal neurons subjected to a chemical hypoxia or glutamate in a dose dependent manner. These data suggest that 5-HT, released during ischemia, may have protective effects in the pathophysiology of ischemic brain damage through a direct action on neurons mediated via the inhibitory 5-HT1A receptor subtype. The results obtained from different in vivo and in vitro models indicate that 5-HT1A agonists are promising agents for the treatment of ischemic brain disorders.

Animals↗

PubMed1993

Endothelin-1 exacerbates focal cerebral ischemia without exerting neurotoxic action in vitro.

In a model of focal cerebral ischemia in mice, intracisternal injection of 5 and 10 pmol/mouse endothelin-1 significantly increased the infarcted surface area by 15.5% and by 23.5%, respectively. Endothelin-1 (0.01, 1, and 100 nmol/l) added to the primary neuronal cultures of chick embryo cerebral hemispheres for 1 h and 24 h did not influence the viability of the neurons or the protein content of the cultures. When applied simultaneously with 1 mmol/l sodium cyanide for 30 min, endothelin-1 (0.01, 1, and 100 nM) did not modify the hypoxia-induced changes. The results show that exogenously applied endothelin-1 could exacerbate cerebral ischemia, probably due to its vasoconstrictive properties and not to a direct neurotoxic effect.

Animals↗

PubMed1993

Protective effects of calpain inhibitors against neuronal damage caused by cytotoxic hypoxia in vitro and ischemia in vivo.

The calpains are calcium-dependent intracellular proteases that are activated in a number of pathogenic conditions. We tested the capacities of protease inhibitors, calpain inhibitor I and leupeptin, to protect against the neuronal degeneration caused by cytotoxic hypoxia or transient global cerebral ischemia. Primary neuronal cultures were prepared from embryonic chick telencephalon, and cytotoxic hypoxia was induced by adding 1 mM NaCN to the culture medium for 30 min. Global ischemia was induced in rats by clamping both carotid arteries and lowering the arterial blood pressure to 40 mmHg for 10 min. Both calpain inhibitor I and leupeptin protected neurons against ischemic and hypoxic damage. Neuroprotection was indicated by increased cell viability and protein content in the cultures, and fewer damaged neurons in the hippocampal CA1-subfield. Thus, blockade of proteolysis can protect neurons against cytotoxic and ischemic damage.

Animals↗

PubMed1993

Platelet-activating factor antagonists reduce excitotoxic damage in cultured neurons from embryonic chick telencephalon and protect the rat hippocampus and neocortex from ischemic injury in vivo.

The neuroprotective effects of the platelet-activating factor (PAF) antagonists BN 52020 and BN 52021 were determined in a temperature-controlled model of transient forebrain ischemia in the rat (occlusion of both common carotid arteries combined with lowering of the mean arterial blood pressure to 40 mm Hg for 10 min). After 7 days of recirculation, the ischemic neuronal damage was evaluated histologically within the hippocampus and neocortex. Combined pre- and post-treatment with the PAF antagonists (2 x 25 mg/kg, s.c.) significantly reduced the resulting neuronal damage of the CA1 and CA3 hippocampal subfields and of the occipital and parietal cerebral cortex. The two PAF antagonists were also tested for their neuroprotective activity in primary neuronal cultures isolated from embryonic chick telencephalon. Since an excessive activation of excitatory amino acid receptors is discussed to be of importance for the ischemic brain damage, the cultured neurons were exposed to the excitatory amino acid L-glutamate (1 mM) for a period of 60 min. Twenty hours after the excitotoxic insult, BN 52020- and BN 52021-treated cultures (1-100 microM) showed both a better preserved morphology, as well as a dose-dependent increase in cell viability and protein content compared to the control cultures. Our results demonstrate that the PAF antagonists BN 52020 and BN 52021 have the capacity to protect brain tissue against ischemic neuronal damage independent of hypothermic effects and are also capable of reducing excitotoxic damage of telencephalic neurons from chick embryos cultured in the absence of glial or endothelial cells. We thus propose that PAF plays an important role in the pathophysiology of ischemic/excitotoxic neuronal injury via a direct action on neurons.

Animals↗

PubMed1993

Mechanisms of drug actions against neuronal damage caused by ischemia--an overview.

1. Oxygen and energy deficits induces a cascade of pathological processes which lead to neuronal dysfunction and cell death. 2. The pathogenesis of ischemia-induced neuronal damage includes a disturbed calcium homeostasis, an excessive release of EAA and an enhanced formation of free oxygen radicals. 3. Calcium antagonists inhibit Ca2+ influx into the neuronal cell via VSCC. 4. Glutamate antagonists reduce intracellular Ca2+ concentration by inactivation of NMDA receptor-associated calcium channels (NMDA antagonists) or AMPA/quisqualate receptor-linked sodium channels (non-NMDA antagonists). 5. Furthermore, oxygen radical scavengers can avoid neuronal damage. 6. Agonists of the adenosinergic and serotonergic transmitter systems contribute to neuroprotection by hyperpolarization of the neuronal membrane due to an increase of K+ permeability.

Brain Ischemia↗

PubMed1993

Transforming growth factor-beta 1 prevents glutamate neurotoxicity in rat neocortical cultures and protects mouse neocortex from ischemic injury in vivo.

Transforming growth factor-beta 1 (TGF-beta 1) has been shown to be an injury-related peptide growth factor within the mammalian central nervous system. We tested whether TGF-beta 1 has the capacity to protect rat neocortical neurons against excitotoxic damage in vitro and mouse neocortex against ischemic injury in vivo. After 14 days in vitro, cultured neurons from rat cerebral cortex were exposed to 1 mM L-glutamate in serum-free culture medium. The cultures received TGF-beta 1 immediately after the addition of glutamate. Eighteen hours later, the cell viability of the cultures was determined using trypan blue exclusion. TGF-beta 1 (1-10 ng/ml) significantly reduced the excitotoxic neuronal damage in a concentration-dependent manner. In vivo, male NMRI mice were subjected to a permanent occlusion of the left middle cerebral artery by microbipolar electrocoagulation. After 48 h, the animals received a transcardiac injection of carbon black. The area of ischemia (devoid of carbon) was restricted to the neocortex and its size was determined planimetrically by means of an image-analyzing system. The treatment with TGF-beta 1 (1 microgram/kg i.c.v.) at 6, 4, or 2 h prior to vessel occlusion reduced the area of ischemia by 5.3, 10.0, and 9.6%, respectively. The effect of the treatment with TGF-beta 1 was statistically significant (p < 0.05 by two-way ANOVA). The present in vitro and in vivo data suggest that TGF-beta 1 has the capacity to diminish the deleterious consequences of an excitotoxic or ischemic insult.

Animals↗

PubMed1993

Isoform-specific effects of transforming growth factors-beta on degeneration of primary neuronal cultures induced by cytotoxic hypoxia or glutamate.

The transforming growth factors-beta (TGFs-beta) are multifunctional peptide growth factors that have been localized in neuronal and glial cells of the CNS of mice, rats, and chick embryos. We tested the TGF-beta isoforms 1, 2, and 3 for their protective effects against neuronal degeneration caused by cytotoxic hypoxia or by the excitatory amino acid L-glutamate. A cytotoxic hypoxia was induced in cultured chick embryo telencephalic neurons by adding 1 mM sodium cyanide to the culture medium for a period of 30 min. Treatment with TGF-beta 1 (1-30 ng/ml) led to a statistically significant increase in cell viability, neuronal ATP levels, and protein content of the cultures assessed 72 h after the toxic insult. TGF-beta 3 was able to reduce the cyanide-induced neuronal damage at concentrations of 0.3 and 1 ng/ml, whereas TGF-beta 2 only showed neuroprotective activity at concentrations of 30 and 50 ng/ml. Both pre- and post-treatment with TGF-beta 1 also prevented the degeneration of cultured chick embryo telencephalic neurons that had been exposed to 1 mM L-glutamate in a buffered salt solution for a period of 60 min. Furthermore, TGF-beta 1 (0.3-3 ng/ml), and to a lesser extent TGF-beta 3 (0.1-1 ng/ml), significantly reduced excitotoxic injury of cultured neurons from rat cerebral cortex that had been exposed to serum-free culture medium supplemented with 1 mM L-glutamate. These results demonstrate that the TGFs-beta are able to prevent the degeneration of primary neuronal cultures, which was caused by energy depletion and activation of glutamate receptors, in an isoform-specific manner.

Animals↗

PubMed1993

Cerebral energy metabolism in fetal guinea pigs during moderate maternal hypoxemia at 0.75 of gestation.

There is evidence from fetal sheep near term that cerebral oxygen delivery decreases during a moderate maternal hypoxemia, whereas cerebral oxygen consumption is maintained. However, since in immature fetuses circulatory centralization may be in part ineffective, cerebral concentrations of high-energy phosphates may fall during a moderate maternal hypoxemia due to an insufficient cerebral oxygen supply. On the other hand anaerobic glycolysis may accelerate to prevent the depletion of high-energy phosphates. To determine, whether or not there is an energy failure in the immature fetal brain in this situation, we studied the effects of 60 min moderate maternal hypoxemia on cerebral concentrations of high-energy phosphates and glycolytic intermediates in fetal guinea pigs at 0.75 gestation. Maternal hypoxemia resulted in no change in fetal mixed arterio-venous pH (7.41 +/- 0.05 vs. 7.38 +/- 0.05; n.s.) or PCO2 (39.0 +/- 4.2 vs. 36.4 +/- 5.6 mmHg; n.s.), but in a fall in fetal PO2 (17.3 +/- 2.2 vs. 11.2 +/- 1.9 mmHg; P < 0.01). There was an increase in fetal cerebral concentrations of lactate (1.50 +/- 0.24 vs. 2.82 +/- 0.91 mumol/g; P < 0.01), whereas those of high-energy phosphates remained unchanged. From these results we conclude that during moderate isocapnic hypoxemia cerebral energy metabolism of immature fetal guinea pigs is maintained by an acceleration of the anaerobic glycolytic rate.

Animals↗

PubMed1992

Extract of kava (Piper methysticum) and its methysticin constituents protect brain tissue against ischemic damage in rodents.

The purpose of the present study was to test whether kava extract and its constituents kawain, dihydrokawain, methysticin, dihydromethysticin and yangonin provide protection against ischemic brain damage. To this end, we used a model of focal cerebral ischemia in mice and rats. Ischemia was induced by microbipolar coagulation of the left middle cerebral artery (MCA). To quantify the size of the lesion in mice, the area of the infarct on the brain surface was assessed planimetrically 48 h after MCA occlusion by transcardial perfusion of carbon black. In the rat model infarct volume was determined 48 h after MCA occlusion by planimetric analysis and subsequent integration of the infarct areas on serial coronal slices. Compounds were administered i.p., except the kava extract, which was administered orally. The effects of the kava extract and its constituents were compared with those produced by the typical anticonvulsant, memantine. The kava extract, methysticin and dihydromethysticin produced effects similar to those of the reference substance memantine. The kava extract (150 mg/kg, 1 h before ischemia) diminished the infarct area (P less than 0.05) in mouse brains and the infarct volume (P less than 0.05) in rat brains. Methysticin, dihydromethysticin (both 10 and 30 mg/kg, 15 min before ischemia) and memantine (20 mg/kg, 30 min before ischemia) significantly reduced the infarct area in mouse brains. All other compounds failed to produce a beneficial effect on the infarct area in mouse brains. In conclusion, the kava extract exhibited neuroprotective activity, which was probably mediated by its constituents methysticin and dihydromethysticin.

Animals↗