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Involvement of platelet-activating factor (PAF) in glutamate neurotoxicity in rat neuronal cultures - PubMed

  • ️Wed Jan 01 1997

Involvement of platelet-activating factor (PAF) in glutamate neurotoxicity in rat neuronal cultures

K Nogami et al. Brain Res. 1997.

Abstract

To clarify the role of platelet-activating factor (PAF) in glutamate neurotoxicity, in vitro experiments using primary neuronal cultures were performed. The anti-PAF immunoglobulin-G (aPAF-IgG) and the three PAF receptor antagonists (BN52021, CV6209, and E5880) were tested for their neuroprotective activity in primary neuronal cultures isolated from embryonic rat cerebral cortex. The cultured neurons were exposed to glutamate (1 mM) for 60 min. Twenty-four hours after this exposure, aPAF-IgG demonstrated evidence of protective effects against neuronal damage in a dose-dependent manner. Protective effects also were observed in cultures treated with the three PAF antagonists (P < 0.05 at 1 microg/ml aPAF-IgG, P < 0.01 at 100 microM BN52021, P < 0.05 at 10 nM CV6209 and P < 0.01 at 10 nM E5880). The Fura-2 assay was used to estimate whether low dosages of exogenous PAF affect cultured neurons. The cultured neurons were loaded with Fura-2/AM. After preincubation for 120 min, the Fura-2-loaded neurons were exposed to various concentrations of PAF for 60 min. By measuring the fluorescent intensity of the medium as representing the amount of Fura-2 released from damaged neurons, we detected an increased release of Fura-2, even at low doses of PAF (P < 0.01 at 10 nM PAF). We further studied PAF production by neurons in response to glutamate. The level of PAF measured in the medium exposed to glutamate was significantly higher than the level in the medium unexposed to glutamate (P < 0.05). Our results suggest an important role of PAF in glutamate neurotoxicity.

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