Enhanced relative expression of glutamate receptor 1 flip AMPA receptor subunits in hippocampal astrocytes of epilepsy patients with Ammon's horn sclerosis.
Astrocytes express ionotropic glutamate receptors (GluRs), and recent evidence suggests that these receptors contribute to direct signaling between neurons and glial cells in vivo. Here, we have used functional and molecular analyses to investigate receptor properties in astrocytes of human hippocampus resected from patients with pharmacoresistant temporal lobe epilepsy (TLE). Histopathological analysis allowed us to distinguish two forms of epilepsy: Ammon's horn sclerosis (AHS) and lesion-associated TLE. Human hippocampal astrocytes selectively expressed the AMPA subtype of ionotropic glutamate receptors. Single-cell RT-PCR found preferential expression of the subunits GluR1 and GluR2 in human astrocytes, and the expression patterns were similar in patients with AHS and lesion-associated epilepsy. The AMPA receptor-specific modulators, cyclothiazide (CTZ) and 4-[2-(phenylsulfonylamino)ethylthio]-2,6-difluoro-phenoxyacetamide (PEPA), were used to investigate splice variant expression. Astrocytes of sclerotic specimens displayed a slower dissociation of CTZ from the receptor and a lower ratio of current potentiation by PEPA to potentiation by CTZ, suggesting enhanced expression of flip receptor variants in AHS versus lesion-associated epilepsy. Real-time PCR and restriction analysis substantiated this presumption by identifying elevated flip-to-flop mRNA ratios of GluR1 in single astrocytes of AHS specimens. These findings imply that in AHS, glutamate may lead to prolonged depolarization of astrocytes, thereby facilitating the generation or spread of seizure activity.
Experimental Neurobiology, Department of Neurosurgery, University of Bonn, 53105 Bonn, Germany. Gerald.Seifert@ukb.uni-bonn.de
Gerald
G
Seifert
Kerstin
K
Hüttmann
Johannes
J
Schramm
Christian
C
Steinhäuser
D017398:Alternative Splicing; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D001581:Benzothiadiazines / Q000494:pharmacology; D002469:Cell Separation; D004827:Epilepsy / Q000503:physiopathology*; D006624:Hippocampus / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D018408:Patch-Clamp Techniques; D010642:Phenoxyacetates / Q000494:pharmacology; D021122:Protein Subunits / Q000378:metabolism; D018091:Receptors, AMPA / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D017470:Receptors, Glutamate / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012598:Sclerosis
D001581:Benzothiadiazines; D010642:Phenoxyacetates; D021122:Protein Subunits; D018091:Receptors, AMPA; D017470:Receptors, Glutamate; C108189:4-(2-(phenylsulfonylamino)ethylthio)-2,6-difluorophenoxyacetamide; C104722:glutamate receptor ionotropic, AMPA 2; C004639:cyclothiazide