Experimental protocol for studying delayed effects of in vitro ischemia on neurotransmitter release from brain slices.
The mechanisms by which ischemic injury leads to delayed neuronal death are not completely understood. Notably, no data are so far available on the modifications in neurosecretory responses evoked by a period of ischemia. Superfused brain slices represent a useful preparation in studying the effects of in vitro ischemia on neurotransmitter release. Using this experimental model we describe a protocol which allows to study not only the immediate effects of an ischemic insult, but also, more interestingly, its delayed (1 h) effects on the release of different neurotransmitters. A first pulse (S1) of 50 mM KCl was applied at the 60th min of perfusion and a second one was applied at the 210th min (S2). In vitro ischemia was performed from the 120th to the 150th min, during the inclusive period between the two depolarizing stimuli. The delayed effects of the ischemic treatment on slice response to KCl were calculated as S2/S1 ratio. This protocol allows to study neurotransmitter release mechanisms associated with postischemic neuronal death. Moreover it will be useful in the evaluation of the neuroprotective potential of new drugs.
Department of Experimental and Clinical Medicine, Section of Pharmacology, University of Ferrara, Via Fossato di Mortara 17, 44100, Ferrara, Italy.
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Sbrenna
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Calò
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Siniscalchi
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Badini
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Bianchi
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Beani
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002545:Brain Ischemia / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D018377:Neurotransmitter Agents / Q000378:metabolism*; D013997:Time Factors
D018377:Neurotransmitter Agents