title: Delta subunit susceptibility variants E177A and R220H associated with complex epilepsy alter channel gating and surface expression of alpha4beta2delta GABAA receptors.
abstract: Most human idiopathic generalized epilepsies (IGEs) are polygenic, but virtually nothing is known of the molecular basis for any of the complex epilepsies. Recently, two GABAA receptor delta subunit variants (E177A, R220H) were proposed as susceptibility alleles for generalized epilepsy with febrile seizures plus and juvenile myoclonic epilepsy. In human embryonic kidney 293T cells, recombinant halpha1beta2delta(E177A) and halpha1beta2delta(R220H) receptor currents were reduced, but the basis for the current reduction was not determined. We examined the mechanistic basis for the current reduction produced by these variants using the halpha4beta2delta receptor, an isoform more physiologically relevant and linked to epileptogenesis, by characterizing the effects of these variants on receptor cell surface expression and single-channel gating properties. Expression of variant alpha4beta2delta(R220H) receptors resulted in a decrease in surface receptor proteins, and a smaller, but significant, reduction was observed for variant alpha4beta2delta(E177A) receptors. For both variants, no significant alterations of surface expression were observed for mixed population of wild-type and variant receptors. The mean open durations of alpha4beta2delta(E177A) and alpha4beta2delta(R220H) receptor single-channel currents were both significantly decreased compared to wild-type receptors. These data suggest that both delta(E177A) and delta(R220H) variants may result in disinhibition in IGEs by similar cellular and molecular mechanisms, and in heterozygously affected individuals, a reduction in channel open duration of delta subunit-containing GABAA receptors may be the major contributor to the epilepsy phenotypes.
authors.affiliation: Department of Neurology, Vanderbilt University Medical Center, Nashville, Tennessee 37212, USA.
authors.forename: Hua-Jun
authors.initials: HJ
authors.lastname: Feng
authors.forename: Jing-Qiong
authors.initials: JQ
authors.lastname: Kang
authors.forename: Luyan
authors.initials: L
authors.lastname: Song
authors.forename: Leanne
authors.initials: L
authors.lastname: Dibbens
authors.forename: John
authors.initials: J
authors.lastname: Mulley
authors.forename: Robert L
authors.initials: RL
authors.lastname: Macdonald
mesh_terms: D019943:Amino Acid Substitution; D002460:Cell Line; D004553:Electric Conductivity; D004829:Epilepsy, Generalized / Q000235:genetics*; D020022:Genetic Predisposition to Disease; D006801:Humans; D015640:Ion Channel Gating / Q000235:genetics*; D007700:Kinetics; D018408:Patch-Clamp Techniques; D011963:Receptors, GABA-A / Q000235:genetics* / Q000378:metabolism*
chemical_list: C546260:GABRA4 protein, human; C495710:GABRB2 protein, human; D011963:Receptors, GABA-A