Effects of GABAA Receptor α3 Subunit Epilepsy Mutations on Inhibitory Synaptic Signaling

Syed，Parnayan1; Durisic，Nela1; Harvey，Robert J.2,3; Sah，Pankaj1,4; Lynch，Joseph W.1

2020-11-20

10.3389/fnmol.2020.602559

Frontiers in Molecular Neuroscience   影响因子和分区

1662-5099

Missense mutations T166M, Q242L, T336M, and Y474C in the GABA receptor (GABAR) α3 subunit gene are associated with epileptic seizures, dysmorphic features, intellectual disability, and developmental delay. When incorporated into GABARs expressed in oocytes, all mutations are known to reduce GABA-evoked whole-cell currents. However, their impact on the properties of inhibitory synaptic currents (IPSCs) is unknown, largely because it is difficult to establish, much less control, the stoichiometry of GABAR expressed in native neuronal synapses. To circumvent this problem, we employed a HEK293 cell-neuron co-culture expression system that permits the recording of IPSCs mediated by a pure population of GABARs with a defined stoichiometry. We first demonstrated that IPSCs mediated by α3-containing GABARs (α3β3γ2) decay significantly slower than those mediated by α1-containing isoforms (α1β2γ2 or α1β3γ2). GABAR α3 mutations did not affect IPSC peak amplitudes or 10–90% rise times, but three of the mutations affected IPSC decay. T336M significantly accelerated the IPSC decay rate whereas T166M and Y474C had the opposite effect. The acceleration of IPSC decay kinetics caused by the T366M mutation was returned to wild-type-like values by the anti-epileptic medication, midazolam. Quantification experiments in HEK293 cells revealed a significant reduction in cell-surface expression for all mutants, in agreement with previous oocyte data. Taken together, our results show that impaired surface expression and altered IPSC decay rates could both be significant factors underlying the pathologies associated with these mutations.

epilepsy GABAA receptor GABRA3 IPSC missense mutation α3 subunit

SCI

英语

其他

National Health and Medical Research Council[1058542][1147600][1156673] ; Australian Research Council[CE140100007]

Neurosciences & Neurology

Neurosciences

WOS:000596049900001

FRONTIERS MEDIA SA

2-s2.0-85097249855

Scopus

1.Queensland Brain Institute,The University of Queensland,Brisbane,Australia
2.School of Health and Behavioural Sciences,University of the Sunshine Coast,Maroochydore,Australia
3.Sunshine Coast Health Institute,Birtinya,Australia
4.Department of Biology,Joint Center for Neuroscience and Neural Engineering,Southern University of Science and Technology,Shenzhen,China

Syed，Parnayan,Durisic，Nela,Harvey，Robert J.,et al. Effects of GABAA Receptor α3 Subunit Epilepsy Mutations on Inhibitory Synaptic Signaling[J]. Frontiers in Molecular Neuroscience,2020,13.

Syed，Parnayan,Durisic，Nela,Harvey，Robert J.,Sah，Pankaj,&Lynch，Joseph W..(2020).Effects of GABAA Receptor α3 Subunit Epilepsy Mutations on Inhibitory Synaptic Signaling.Frontiers in Molecular Neuroscience,13.

Syed，Parnayan,et al."Effects of GABAA Receptor α3 Subunit Epilepsy Mutations on Inhibitory Synaptic Signaling".Frontiers in Molecular Neuroscience 13(2020).