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Neuroprotection Promoted by Guanosine Depends on Glutamine Synthetase and Glutamate Transporters Activity in Hippocampal Slices Subjected to Oxygen/Glucose Deprivation

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Abstract

Guanosine (GUO) has been shown to act as a neuroprotective agent against glutamatergic excitotoxicity by increasing glutamate uptake and decreasing its release. In this study, a putative effect of GUO action on glutamate transporters activity modulation was assessed in hippocampal slices subjected to oxygen and glucose deprivation (OGD), an in vitro model of brain ischemia. Slices subjected to OGD showed increased excitatory amino acids release (measured by d-[3H]aspartate release) that was prevented in the presence of GUO (100 µM). The glutamate transporter blockers, DL-TBOA (10 µM), DHK (100 µM, selective inhibitor of GLT-1), and sulfasalazine (SAS, 250 µM, Xc− system inhibitor) decreased OGD-induced d-aspartate release. Interestingly, DHK or DL-TBOA blocked the decrease in glutamate release induced by GUO, whereas SAS did not modify the GUO effect. GUO protected hippocampal slices from cellular damage by modulation of glutamate transporters, however selective blockade of GLT-1 or Xc- system only did not affect this protective action of GUO. OGD decreased hippocampal glutamine synthetase (GS) activity and GUO recovered GS activity to control levels without altering the kinetic parameters of GS activity, thus suggesting GUO does not directly interact with GS. Additionally, the pharmacological inhibition of GS activity with methionine sulfoximine abolished the effect of GUO in reducing d-aspartate release and cellular damage evoked by OGD. Altogether, results in hippocampal slices subjected to OGD show that GUO counteracts the release of excitatory amino acids, stimulates the activity of GS, and decreases the cellular damage by modulation of glutamate transporters activity.

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Abbreviations

DHK:

Dihydrokainate

DL-TBOA:

DL-threo-β-benzyloxyaspartic acid

GS:

Glutamine synthetase

GUO:

Guanosine

HBSS:

Hank’s balanced salt solution

KRB:

Krebs-Ringer bicarbonate buffer

MSO:

Methionine sulfoximine

OGD:

Oxygen/glucose deprivation

SAS:

Sulfasalazine

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Acknowledgments

Research supported by grants from the Brazilian funding agencies CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico)—Projects: IBN-Net # 01.06.0842-00 and INCT (Instituto Nacional de Ciência e Tecnologia) for Excitotoxicity and Neuroprotection and FAPESC (Pronex/NENASC) to C.I.T. C.I.T. is recipient of CNPq productivity fellowship and T. D.-C. is recipient of CAPES-PVE (052/2012) post-doctoral scholarship.

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Authors and Affiliations

  1. Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Trindade, Florianópolis, SC, 88040-900, Brazil

    Tharine Dal-Cim, Wagner C. Martins, Daniel T. Thomaz, Victor Coelho, Gabriela Godoy Poluceno, Débora Lanznaster & Carla I. Tasca

  2. Programa de pós-graduação em Neurociências, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil

    Tharine Dal-Cim, Wagner C. Martins, Débora Lanznaster & Carla I. Tasca

  3. Programa de pós-graduação em Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil

    Daniel T. Thomaz & Carla I. Tasca

  4. Departamento de Ciências Básicas em Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso, Cuiabá, MT, Brazil

    Samuel Vandresen-Filho

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  1. Tharine Dal-Cim
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  2. Wagner C. Martins
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Correspondence to Carla I. Tasca.

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Dal-Cim, T., Martins, W.C., Thomaz, D.T. et al. Neuroprotection Promoted by Guanosine Depends on Glutamine Synthetase and Glutamate Transporters Activity in Hippocampal Slices Subjected to Oxygen/Glucose Deprivation. Neurotox Res 29, 460–468 (2016). https://doi.org/10.1007/s12640-015-9595-z

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