Delayed protection by MK-801 and tetrodotoxin in a rat organotypic hippocampal culture model of ischemia.

Vornov, James J.; Tasker, Robert C.; Coyle, Joseph T.

doi:10.1161/01.str.25.2.457

Cited by 95 publications

(43 citation statements)

References 33 publications

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“…In these experiments, ischemialike conditions are induced by a combination of OGD achieved by an anoxic atmosphere in combination with a glucose-free medium in a manner similar to that described for dissociated cell cultures (Goldberg and Choi, 1993;Rothman 1983), or by inhibition of oxidative phosphorylation using cyanide in combination with 2-deoxyglucose (Bernaudin et al, 1998;Vornov et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

Mouse Hippocampal Organotypic Tissue Cultures Exposed to In Vitro “Ischemia” Show Selective and Delayed CA1 Damage that is Aggravated by Glucose

Rytter

Cronberg

Asztély

et al. 2003

J Cereb Blood Flow Metab

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Summary:Oxygen and glucose deprivation (OGD) in cell cultures is generally studied in a medium, such as artificial cerebrospinal fluid (CSF), with an ion composition similar to that of the extracellular fluid of the normal brain (2 to 4 mmol/L K + , 2 to 3 mmol/L Ca 2+ ; pH 7.4). Because the distribution of ions across cell membranes dramatically shifts during ischemia, the authors exposed mouse organotypic hippocampal tissue cultures to OGD in a medium, an ischemic cerebrospinal fluid, with an ion composition similar to the extracellular fluid of the brain during ischemia in vivo (70 mmol/L K + , 0.3 mmol/L Ca 2+ ; pH 6.8). In ischemic CSF, OGD induced a selective and delayed cell death in the CA1 region, as assessed by propidium iodide uptake. Cell death was glutamate receptor dependent since blockade of the N-methyl-D-aspartate and ␣-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors mitigated cell damage. Hyperglycemia aggravates ischemic brain damage in vivo, whereas in vitro glucose in artificial CSF prevents oxygen deprivation-induced damage. The authors demonstrate that glucose in ischemic CSF significantly exacerbates cell damage after oxygen deprivation. This new model of in vitro "ischemia" can be useful in future studies of the mechanisms and treatment of ischemic cell death, including studies using genetically modified mice.

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Section: Discussionmentioning

confidence: 99%

Mouse Hippocampal Organotypic Tissue Cultures Exposed to In Vitro “Ischemia” Show Selective and Delayed CA1 Damage that is Aggravated by Glucose

Rytter

Cronberg

Asztély

et al. 2003

J Cereb Blood Flow Metab

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“…excitotoxins [2,28,34,38], hypoxia rich, capsule-like zone devoid of MAP2-immunoreactive and hypoglycemia alone or in combination [23, 25,35], processes. This might be a reaction of the tissue to silicon nitric oxide [1] and trimethyltin [19,20].…”

Section: Discussionmentioning

confidence: 99%

Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures

et al. 2001

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In this study we examined the passive biocompatibility of a three-dimensional microelectrode array (MEA), designed to be coupled to organotypic brain slice cultures for multisite recording of electrophysiological signals. Hippocampal (and corticostriatal) brain slices from 1-week-old (and newborn) rats were grown for 4-8 weeks on the perforated silicon chips with silicon nitride surfaces and 40 mm sized holes and compared with corresponding tissue slices grown on conventional semiporous membranes. In terms of preservation of the basic cellular and connective organization, as visualized by Nissl staining, Timm sulphide silver-staining, microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP) immunostaining, the slice cultures grown on chips did not differ from conventionally grown slice cultures. Neither were there any signs of astrogliosis or neurodegeneration around the upper recording part of the 47-mm-high platinum-tip electrodes. Slice cultures grown on a separate set of chips with platinum instead of silicon nitride surfaces also displayed normal MAP2 and GFAP immunostaining. The width of the GFAP-rich zone (glia limitans) at the bottom surface of the slice cultures was the same (|20 mm) in cultures grown on chips with silicon nitride and platinum surfaces and on conventional insert membranes. The slice cultures grown on chips maintained a normal, subfield differentiated susceptibility to the glutamate receptor agonist N-methyl-D-aspartate (NMDA) and the neurotoxin trimethyltin (TMT), as demonstrated by the cellular uptake of propidium iodide (PI), which was used as a reproducible and quantifiable marker for neuronal degeneration. We conclude that organotypic brain slice cultures can grow on silicon-based three-dimensional microelectrode arrays and develop normally with display of normal subfield differentiated susceptibilities to known excito-and neurotoxins. From this it is anticipated that the set-up, designed for recording of electrophysiological parameters, can be used for long-term studies of defined neuronal networks and provide valuable information on both normal, neurotoxicological and neuropathological conditions. Theme: Disorders of the nervous systemTopic: Neurotoxicity

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“…Regional patterns of neuronal injury in the organotypic cultures were observed by performing experiments in the presence of propidium iodide. After membrane injury, the dye enters cells, binds to nucleic acids, and accumulates, rendering the cell brightly fluorescent (27). The CA1 neuronal subfield was clearly visible in a bright field image.…”

Section: Preparation Of Primary Neuronal Culturesmentioning

confidence: 99%

Galanin acts as a neuroprotective factor to the hippocampus

Elliott-Hunt

Marsh

Bacon

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

131

105

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The expression of the neuropeptide galanin is markedly up-regulated in many areas of the central and peripheral nervous system after injury. We have recently demonstrated that peripheral sensory neurons depend on galanin for neurite extension after injury, mediated by activation of the second galanin receptor subtype (GALR2). We therefore hypothesized that galanin might also act in a similar manner in the CNS, reducing cell death in hippocampal models of excitotoxicity. Here we report that galanin acts an endogenous neuroprotective factor to the hippocampus in a number of in vivo and in vitro models of injury. Kainate-induced hippocampal cell death was greater in both the CA1 and CA3 regions of galanin knockout animals than in WT controls. Similarly, exposure to glutamate or staurosporine induced significantly more neuronal cell death in galanin knockout organotypic and dispersed primary hippocampal cultures than in WT controls. Conversely, less cell death was observed in the hippocampus of galanin overexpressing transgenic animals after kainate injection and in organotypic cultures after exposure to staurosporine. Further, exogenous galanin or the previously described high-affinity GALR2 agonist, both reduced cell death when coadministered with glutamate or staurosporine in WT cultures. These results demonstrate that galanin acts an endogenous neuroprotective factor to the hippocampus and imply that a galanin agonist might have therapeutic uses in some forms of brain injury.T he 29-aa neuropeptide galanin (1) is widely expressed in both the central and peripheral nervous system and has strong inhibitory actions on synaptic transmission by reducing the release of a number of classical neurotransmitters (2-7). These inhibitory actions result in a diverse range of physiological effects including, an impairment of working memory (8) and long-term potentiation (9); a reduction in hippocampal excitability with a decreased predisposition to seizure activity (10); and a marked inhibition of nociceptive responses in the intact animal and after nerve injury (11). These neuromodulatory actions of galanin have long been regarded as the principal role played by the peptide in the nervous system. However, there is now a large body of evidence to indicate that injury to many of these neuronal systems markedly induces the expression of galanin at both the mRNA and peptide levels. Examples of such lesion studies include the up-regulation of galanin in (i) the dorsal root ganglion after peripheral nerve axotomy (12), (ii) magnocellular secretory neurons of the hypothalamus after hypophysectomy (13), (iii) the dorsal raphe and thalamus after removal of the frontoparietal cortex (decortication) (14), (iv) the molecular layer of the hippocampus after an entorhinal cortex lesion (15), and (v) the medial septum and vertical limb diagonal-band after a fimbria fornix bundle transection (16). These studies have led a number of investigators to speculate that galanin might play a trophic role in addition to its classical neuromodulatory ...

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Delayed protection by MK-801 and tetrodotoxin in a rat organotypic hippocampal culture model of ischemia.

Cited by 95 publications

References 33 publications

Mouse Hippocampal Organotypic Tissue Cultures Exposed to In Vitro “Ischemia” Show Selective and Delayed CA1 Damage that is Aggravated by Glucose

Mouse Hippocampal Organotypic Tissue Cultures Exposed to In Vitro “Ischemia” Show Selective and Delayed CA1 Damage that is Aggravated by Glucose

Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures

Galanin acts as a neuroprotective factor to the hippocampus

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