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Time-dependent modulation of AMPA receptor phosphorylation and mRNA expression of NMDA receptors and glial glutamate transporters in the rat hippocampus and cerebral cortex in a pilocarpine model of epilepsy

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Abstract

The pilocarpine model in rodents reproduces the main features of mesial temporal lobe epilepsy related to hippocampus sclerosis (MTLE-HS) in humans. It has been demonstrated in this model that the phosphorylation of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor GluR1 subunit is increased 1 h after pilocarpine treatment. Moreover, alterations in the levels of glutamate transporters have been associated with chronic epilepsy in humans. Despite these studies, the profile of these changes has not yet been addressed. We analyzed the protein content and phosphorylation profile of the AMPA receptor GluR1 subunit by western blotting. We also used quantitative real-time polymerase chain reaction to analyze the expression of glial glutamate transporters and the N-methyl-d-aspartate receptor NR1 subunit in the hippocampus (Hip) and cerebral cortex (Ctx) at different time points after pilocarpine-induced status epilepticus (Pilo-SE) in male adult Wistar rats. Biochemical analysis was performed in the Hip and Ctx at 1, 3, 12 h (acute period), 5 days (latent period), and 50 days (chronic period) after Pilo-SE. Key findings include an increase in the phosphorylation of GluR1-Ser845 in the Ctx and GluR1-Ser831 in the Hip at different times during the acute period, and a decrease in the total content of the GluR1 subunit in the Ctx in the latent period. There was a down-regulation of the mRNA expression and protein levels of EAAT1 and EAAT2, and a decrease of the NR1 mRNA expression, in the Ctx during the latent period. Notably, during the chronic period, the EAAT2 mRNA expression and protein levels decreased while the NR1 mRNA levels increased in the Hip. Taken together, our findings suggest a time- and structure-dependent imbalance of glutamatergic transmission in response to Pilo-SE, which might be associated with either epileptogenesis or the seizure threshold in MTLE-HS.

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Acknowledgments

This work was supported by the National Council for Scientific and Technological Development (CNPq) Brazil; Coordination for the Training and Improvement of Higher Education Personnel (CAPES); Santa Catarina Research Foundation (FAPESC/PRONEX Program-NENASC project), INCT-National Institute of Science and Technology for Excitotoxicity and Neuroprotection and IBN.Net/CNPq. R.B.L. and R.W. are recipients of CNPq fellowships.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

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

    Mark William Lopes, Fabiano Mendes de Cordova & Rodrigo Bainy Leal

  2. Centro de Neurociências Aplicadas (CeNAp), HU, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil

    Flávia Mahatma Schneider Soares, Nelson de Mello, Jean Costa Nunes & Roger Walz

  3. Centro de Ciências Agrárias e Biológicas, Universidade Estadual Vale do Acaraú, Fortaleza, CE, Brazil

    Aurilene Gomes Cajado, Daniel de Brito & Rodrigo Maranguape Silva da Cunha

  4. Escola de Medicina Veterinária e Zootecnia, Universidade Federal do Tocantins, Araguaína, TO, Brazil

    Fabiano Mendes de Cordova

  5. Departamento de Clínica Médica, Hospital Universitário (HU), Centro de Ciências da Saúde, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil

    Roger Walz

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  1. Mark William Lopes
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  2. Flávia Mahatma Schneider Soares
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Correspondence to Rodrigo Bainy Leal.

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Lopes, M.W., Soares, F.M.S., de Mello, N. et al. Time-dependent modulation of AMPA receptor phosphorylation and mRNA expression of NMDA receptors and glial glutamate transporters in the rat hippocampus and cerebral cortex in a pilocarpine model of epilepsy. Exp Brain Res 226, 153–163 (2013). https://doi.org/10.1007/s00221-013-3421-8

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  • DOI: https://doi.org/10.1007/s00221-013-3421-8

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