Glutamate induced neonatal excitotoxicity modifies the expression level of EAAT1 (GLAST) and EAAT2 (GLT-1) proteins in various brain regions of the adult rat

Neurosci Lett. 2020 Sep 14:735:135237. doi: 10.1016/j.neulet.2020.135237. Epub 2020 Jul 6.

Abstract

Glutamate-mediated excitatory synaptic signalling is primarily controlled by excitatory amino acid transporters (EAATs), such as EAAT1 and EAAT2, which are located mostly on astrocytes and, together, uptake more than 95 % of extracellular glutamate. Alterations in the functional expression levels of EAATs can lead to excessive extracellular glutamate accumulation, potentially triggering excitotoxicity and seizures, among other neurological disorders. Excitotoxicity induced in early developmental stages can lead to lasting changes in several neurotransmission systems, including the glutamatergic system, which could make the brain more susceptible to a second insult. In this study, the expression levels of EAAT1 (GLAST) and EAAT2 (GLT-1) proteins were assessed in the cerebral motor cortex (CMC), striatum, hippocampus and entorhinal cortex (EC) of male adult rats following the neonatal excitotoxic process triggered by monosodium glutamate (MSG)-treatment (4 g/kg of body weight at postnatal days 1,3,5 and 7, subcutaneously). Western blot analysis showed that neonatal MSG-treatment decreased EAAT1 expression levels in the CMC, striatum and hippocampus, while EAAT2 levels were increased in the striatum and EC and decreased in the CMC. Immunofluorescence staining confirmed the changes in EAAT1 and EAAT2 expression induced by neonatal MSG-treatment, which were accompanied by an increase in the glial fibrillary acidic protein (GFAP) immunofluorescence signalthat was particularly significant in the hippocampus. Our results show that a neonatal excitotoxic processes can induce lasting changes in the expression levels of EAAT1 and EAAT2 proteins and suggest that although astrogliosis occurs, glutamate uptake could be deficient, particularly in the CMC and hippocampus.

Keywords: Astrogliosis; EAAT1; EAAT2; Excitotoxicity; Monosodium glutamate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Age Factors
  • Animals
  • Animals, Newborn
  • Brain / drug effects
  • Brain / growth & development*
  • Brain / metabolism*
  • Excitatory Amino Acid Transporter 1 / antagonists & inhibitors
  • Excitatory Amino Acid Transporter 1 / biosynthesis*
  • Excitatory Amino Acid Transporter 1 / genetics
  • Excitatory Amino Acid Transporter 2 / antagonists & inhibitors
  • Excitatory Amino Acid Transporter 2 / biosynthesis*
  • Excitatory Amino Acid Transporter 2 / genetics
  • Gene Expression
  • Glutamic Acid / toxicity
  • Male
  • Rats
  • Rats, Wistar
  • Sodium Glutamate / toxicity*

Substances

  • Excitatory Amino Acid Transporter 1
  • Excitatory Amino Acid Transporter 2
  • Slc1a2 protein, rat
  • Slc1a3 protein, rat
  • Glutamic Acid
  • Sodium Glutamate