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- Moroni Ramona Frida RF Epilepsy Unit, Fondazione I.R.C.C.S. Istituto Neurologico "C. Besta", via Celoria 11, 20133 Milan, Italy. Electronic address: ramona.moroni@istitut, Regondi Maria Cristina MC Epilepsy Unit, Fondazione I.R.C.C.S. Istituto Neurologico "C. Besta", via Celoria 11, 20133 Milan, Italy. Electronic address: cristina.regondi@i, Marco de Curtis, Carolina Frassoni, and Laura Librizzi.
- Epilepsy Unit, Fondazione I.R.C.C.S. Istituto Neurologico "C. Besta", via Celoria 11, 20133 Milan, Italy. Electronic address: ramona.moroni@istituto-besta.it.
- Neuroscience. 2020 Aug 10; 441: 65-76.
AbstractAstrocyte dysfunction, and in particular impaired extracellular potassium spatial buffering, has been postulated to have a potential role in seizure susceptibility and ictogenesis. Inwardly rectifying potassium (Kir) channels, and specifically KIR4.1, have a predominant role in K+ homeostasis and their involvement in neuronal excitability control have been hypothesized. To avoid the severe side effects observed in Kir4.1 cKO, we studied the effects of Kir4.1 down-regulation in cortical astrocytes by using Kir4.1 RNA interference (RNAi) technique combined with in utero electroporation (IUE) at E16 and a piggyBac transposon system. Kir4.1 down-regulation was confirmed by immunohistochemistry and field fraction analysis. To investigate if Kir4.1 silencing affects 4AP-induced seizure threshold and extracellular potassium homeostasis, simultaneous in vitro field potential and extracellular K+ recordings were performed on somatosensory cortex slices obtained from rats electroporated with a piggyBac-Kir4.1-shRNA (Kir4.1-) and scrambled shRNA (Kir4.1Sc). Electrophysiological data revealed no significant differences in terms of seizure onset and seizure-induced extracellular K+ changes between Kir4.1- and Kir4.1Sc rats. Intriguingly, immunohistochemical analysis performed on slices studied with electrophysiology revealed a reduced number of neurons generated from radial glial cells in Kir4.1- rats. We conclude that focal down-regulation of Kir4.1 channel in cortical astrocytes by Kir4.1 RNAi technique combined with IUE is not effective in altering potassium homeostasis and seizure susceptibility. This technique revealed a possible role of Kir4.1 during corticogenesis.Copyright © 2020 IBRO. Published by Elsevier Ltd. All rights reserved.
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