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Energy Substrates Protect Hippocampus Against Endogenous Glutamate-Mediated Neurodegeneration in Awake Rats

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Abstract

Excitotoxicity due to excessive glutamatergic neurotransmission is a well-studied phenomenon that has been related to the mechanisms of neuronal death occurring in some disorders of the CNS. We have previously shown that the intrahippocampal perfusion by microdialysis of 4-aminopyridine (4-AP) in rats stimulates endogenous glutamate release from nerve endings and this results in excitotoxic effects such as immediate seizures and delayed neuronal death, due to the overactivation of N-methyl-d-aspartate (NMDA) receptors. To study whether mitochondrial energy dysfunction and oxidative stress could be involved in this 4-AP-induced excitotoxicity, we evaluated in awake rats the protective effect of several energy substrates and antioxidant compounds, using microdialysis, electroencephalographic (EEG) recording and histological analysis. The 4-AP-induced behavioral and EEG seizures, which progressed to status epilepticus in about 30 min, were prevented by the NMDA receptor antagonist MK-801, whereas acetoacetate, dl- and l-β-hydroxybutyrate did not protect against seizures but increased the latency to the onset of status epilepticus; pyruvate, α-ketoglutarate and glutathione ethyl ester did not show any protective effect. 4-AP also produced nearly complete loss of pyramidal neurons in CA1 and CA3 regions of the ipsilateral hippocampus 24 h after the experiment. MK-801 totally prevented this neuronal death and the energy substrates tested protected by about 50 %, whereas the antioxidants showed only a weak protection. We conclude that ketone bodies possess weak anticonvulsant effects and that energy metabolism impairment plays a more important role than oxidative stress in the delayed hippocampal neurodegeneration resulting from the excitotoxic action of 4-AP mediated by endogenous glutamate.

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Abbreviations

4-AP:

4-Aminopyridine

ACAC:

Acetoacetate

dl-βHB:

dl-β-Hydroxybutyrate

EEG:

Electroencephalogram

GEE:

Glutathione ethyl ester

GSH:

Reduced glutathione

KB:

Ketone bodies

NMDA:

N-Methyl-d-aspartate

Pyr:

Pyruvate

αKG:

α-Ketoglutarate

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Acknowledgments

This work was supported by Consejo Nacional de Ciencia y Tecnología, México (CONACYT, project 128229) and DGAPA, UNAM (IN201013). C.N. is recipient of a scholarship from CONACYT. This work is part of the Ph.D. thesis of C.N.

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  1. División de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, AP 70-253, 04510, Mexico, DF, Mexico

    Citlalli Netzahualcoyotzi & Ricardo Tapia

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  1. Citlalli Netzahualcoyotzi
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  2. Ricardo Tapia
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Correspondence to Ricardo Tapia.

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Netzahualcoyotzi, C., Tapia, R. Energy Substrates Protect Hippocampus Against Endogenous Glutamate-Mediated Neurodegeneration in Awake Rats. Neurochem Res 39, 1346–1354 (2014). https://doi.org/10.1007/s11064-014-1318-y

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  • DOI: https://doi.org/10.1007/s11064-014-1318-y

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