Time-dependent modulation of AMPA receptor phosphorylation and mRNA expression of NMDA receptors and glial glutamate tra
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RESEARCH ARTICLE
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 Mark William Lopes · Flávia Mahatma Schneider Soares · Nelson de Mello · Jean Costa Nunes · Aurilene Gomes Cajado · Daniel de Brito · Fabiano Mendes de Cordova · Rodrigo Maranguape Silva da Cunha · Roger Walz · Rodrigo Bainy Leal Received: 13 June 2012 / Accepted: 15 January 2013 / Published online: 8 February 2013 © Springer-Verlag Berlin Heidelberg 2013
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
M. W. Lopes · F. M. de Cordova · R. B. Leal (*) Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Florianópolis, SC 88040-900, Brazil e-mail: [email protected] F. M. S. Soares · N. de Mello · J. C. Nunes · R. Walz Centro de Neurociências Aplicadas (CeNAp), HU, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil A. G. Cajado · D. de Brito · R. M. S. da Cunha Centro de Ciências Agrárias e Biológicas, Universidade Estadual Vale do Acaraú, Fortaleza, CE, Brazil F. M. de Cordova Escola de Medicina Veterinária e Zootecnia, Universidade Federal do Tocantins, Araguaína, TO, Brazil R. Walz Departamento de Clínica Médica, Hospital Universitário (HU), Centro de Ciências da Saúde, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil
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 NR
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