Profile of phosphoprotein labelling in organotypic slice cultures of rat hippocampus

Tavares, Aline; Cimarosti, Helena; Valentim, Lauren Martins; Salbego, Christianne Gazzana

doi:10.1097/00001756-200108280-00023

Cited by 13 publications

(4 citation statements)

References 22 publications

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“…The response of hippocampal slice cultures to ischemic conditions induced by OGD is similar to that displayed by animals in the situation in vivo (Tavares et al ., 2001), suggesting the suitability of these cultures for the investigation of direct neuronal effects of a drug on ischemic injury in the absence of blood circulation and infiltrating inflammatory cells.…”

Section: Discussionsupporting

confidence: 53%

A synthetic NCAM‐derived peptide, FGL, protects hippocampal neurons from ischemic insult both in vitro and in vivo

Skibo

Lushnikova

Воронин

et al. 2005

Eur J of Neuroscience

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There is a major unmet need for development of innovative strategies for neuroprotection against ischemic brain injury. Here we show that FGL, a neural cell adhesion molecule (NCAM)-derived peptide binding to and inducing phosphorylation of the fibroblast growth factor receptor (FGFR), acts neuroprotectively after an ischemic insult both in vitro and in vivo. The neuroprotective activity of FGL was tested in vitro on dissociated rat hippocampal neurons and hippocampal slice cultures, using a protocol of oxygen-glucose deprivation (OGD). FGL protected hippocampal neurons from damage and maintained or restored their metabolic and presynaptic activity, both if employed as a pretreatment alone to OGD, and if only applied after the insult. In vivo 24 h pretreatment with a single suboccipital injection of FGL significantly protected hippocampal CA1 neurons from death in a transient global ischemia model in the gerbil. We conclude that FGL promotes neuronal survival after ischemic brain injury.

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Section: Discussionsupporting

confidence: 53%

A synthetic NCAM‐derived peptide, FGL, protects hippocampal neurons from ischemic insult both in vitro and in vivo

Skibo

Lushnikova

Воронин

et al. 2005

Eur J of Neuroscience

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“…To model ischemic events organotypic cultures are exposed to oxygen and glucose deprivation (OGD) using an anaerobic chamber. We have found that the response of organotypic cultured tissue to injury induced by OGD is very similar to that shown by animals submitted to transient cerebral ischemia, suggesting the suitability of this model for the study of ischemic lesions and neuroprotective drugs (4,5).…”

Section: Introductionsupporting

confidence: 55%

Estradiol Protects Against Oxygen and Glucose Deprivation in Rat Hippocampal Organotypic Cultures and Activates Akt and Inactivates GSK-3?

et al. 2005

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Here we investigated the neuroprotective effect of 17beta-estradiol in an in vitro model of ischemia. We used organotypic hippocampal slice cultures, acute or chronically treated with 17beta-estradiol (10 nM), and exposed to oxygen and glucose deprivation (OGD). Cellular death was quantified by measuring uptake of propidium iodide (PI), a marker of dead cells. In OGD exposed cultures, treated only with vehicle, about 70% of the CA1 area of hippocampus was labeled with PI, indicating a great percentage of cellular death. When cultures were treated with 17beta-estradiol (acute or chronically), this cellular death was reduced to 15%. This effect was prevented by LY294002 but was not by PD98059. Immunoblotting revealed that both, chronic and acute, treatments with 17beta-estradiol induced the phosphorylation/activation of Akt and the phosphorylation/inactivation of GSK-3beta. Our results show a clear neuroprotective effect of 17beta-estradiol and suggest that this effect could involve PI3-K pathway.

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“…After obtaining the fluorescent images, cultured slices were homogenized in lysis buffer (4% SDS, 2.1 mM EDTA, 50 mM Tris), aliquots were taken for protein determination and b-mercaptoethanol was added to a final concentration of 5% [18]. Samples containing 40 lg of protein were resolved by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).…”

Section: Western Blot Analysismentioning

confidence: 99%

The Effects of Estradiol on Estrogen Receptor and Glutamate Transporter Expression in Organotypic Hippocampal Cultures Exposed to Oxygen--Glucose Deprivation

et al. 2006

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The molecular basis of estrogen-mediated neuroprotection against brain ischemia remains unclear. In the present study, we investigated changes in expression of estrogen receptors (ERs) alpha and beta and excitatory amino acid transporters (EAAT) 1 and 2 in rat organotypic hippocampal slice cultures treated with estradiol and subsequently exposed to oxygen--glucose deprivation (OGD). Pretreatment with 17beta-estradiol (10 nM) for 7 days protected the CA1 area of hippocampus against OGD (60 min), reducing cellular injury by 46% compared to the vehicle control group. Levels of ERalpha protein were significantly reduced by 20% after OGD in both vehicle- and estradiol-treated cultures, whereas ERbeta was significantly up-regulated by 25% in the estradiol-treated cultures. In contrast, EAAT1 and EAAT2 levels were unchanged in response to estradiol treatment in this model of OGD. These findings suggest that estrogen-induced neuroprotection against ischemia might involve regulation of ERbeta and, consequently, of the genes influenced by this receptor.

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Profile of phosphoprotein labelling in organotypic slice cultures of rat hippocampus

Cited by 13 publications

References 22 publications

A synthetic NCAM‐derived peptide, FGL, protects hippocampal neurons from ischemic insult both in vitro and in vivo

A synthetic NCAM‐derived peptide, FGL, protects hippocampal neurons from ischemic insult both in vitro and in vivo

Estradiol Protects Against Oxygen and Glucose Deprivation in Rat Hippocampal Organotypic Cultures and Activates Akt and Inactivates GSK-3?

The Effects of Estradiol on Estrogen Receptor and Glutamate Transporter Expression in Organotypic Hippocampal Cultures Exposed to Oxygen--Glucose Deprivation

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