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Neuroscience. 2004;125(3):803-18.

Response to kainic acid injections: changes in staining for zinc, FOS, cell death and glial response in the rat forebrain.

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  • 1Departament de Biologia Cel.lular, Universitat de Barcelona, Facultat de Biologia, Diagonal 645, ES-08071 Barcelona, Spain. angela@bio.ub.es

Abstract

A pool of zinc is present in synaptic vesicles in a population of glutamatergic neurones. Zinc appears to modulate synaptic transmission and cause neuronal death. The status of vesicular zinc, neuronal death and glial reaction in the rat forebrain was analysed after a systemic injection of kainic acid in order to establish a model for future studies on zinc function. Rats received a systemic injection of kainic acid (10 mg/kg) and were killed 3, 6, 12, 24 and 48 h post-treatment. Timm's method and zinquin staining were used to detect zinc. Immunostaining for Fos-like proteins and staining with Fluoro-Jade B were used to detect cell reaction and degeneration, respectively. Glial fibrillary acidic protein and tomato lectin were used as glial markers. Zinquin staining for zinc rose transitorily in neuronal somata 6 h after injection (not observed at 24-48 h) in the piriform and entorhinal cortices, amygdala and hippocampus. In contrast sulphide/silver staining for zinc showed virtually no rise in cytoplasmic zinc (except in cornus ammonis field 1 of the hippocampus) 6 h after injection, and a decrease (bleaching) in some terminal fields starting 12 h after injection and increasing at 24-48 h. The areas most affected by the zinc bleaching were the olfactory bulb, piriform and entorhinal cortices, endopiriform and amygdaloid nuclei. Transitory Fos immunostaining (within neuronal nuclei) was observed between 3 and 12 h after kainate treatment in many telencephalic areas: olfactory bulb, cortex (piriform, hippocampal and neocortex) and amygdaloid nuclei. This was accompanied by changes in glial markers starting 3 h after injection. Fluoro-Jade B staining in neurones (degeneration) appeared 6 h after treatment and increased later. Degenerating areas generally coincided with those showing Fos immunoreactivity. Zinquin and sulphide/silver methods revealed various pools of zinc after kainate injection: a cytoplasmic pool and a terminal field (or vesicular) pool. Cytoplasmic zinc (zinquin) was coincident, in time and location, with cell degeneration, thus implicating zinc in cell death. This zinc may not have come from presynaptic stores, since no bleaching (sulphide/silver method) was observed 6 h after injection. Future experiments altering zinc pools (e.g. by chelating agents) may elucidate the function of zinc.

PMID:
15099693
[PubMed - indexed for MEDLINE]
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