CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility

Kosuke Okuda; Shizuka Kobayashi; Masahiro Fukaya; Aya Watanabe; Takuto Murakami; Mai Hagiwara; Tempei Sato; Hiroe Ueno; Narumi Ogonuki; Sayaka Komano-Inoue; Hiroyuki Manabe; Masahiro Yamaguchi; Atsuo Ogura; Hiroshi Asahara; Hiroyuki Sakagami; Masashi Mizuguchi; Toshiya Manabe; Teruyuki Tanaka

doi:10.1016/j.nbd.2017.07.002

CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility

Neurobiol Dis. 2017 Oct:106:158-170. doi: 10.1016/j.nbd.2017.07.002. Epub 2017 Jul 6.

Authors

Kosuke Okuda¹, Shizuka Kobayashi², Masahiro Fukaya³, Aya Watanabe¹, Takuto Murakami¹, Mai Hagiwara¹, Tempei Sato⁴, Hiroe Ueno⁴, Narumi Ogonuki⁵, Sayaka Komano-Inoue⁶, Hiroyuki Manabe⁶, Masahiro Yamaguchi⁶, Atsuo Ogura⁵, Hiroshi Asahara⁷, Hiroyuki Sakagami³, Masashi Mizuguchi¹, Toshiya Manabe², Teruyuki Tanaka⁸

Affiliations

¹ Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
² Division of Neuronal Network, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.
³ Department of Anatomy, Kitasato University School of Medicine, Sagamihara 252-0374, Japan.
⁴ Department of Systems Biomedicine, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan.
⁵ Bioresource Engineering Division, RIKEN BioResource Center, Tsukuba 305-0074, Japan.
⁶ Department of Physiology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
⁷ Department of Systems Biomedicine, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan; Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla 92037, USA.
⁸ Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan. Electronic address: tetanaka@m.u-tokyo.ac.jp.

PMID: 28688852
DOI: 10.1016/j.nbd.2017.07.002

Abstract

Mutations in the Cyclin-dependent kinase-like 5 (CDKL5) gene cause severe neurodevelopmental disorders accompanied by intractable epilepsies, i.e. West syndrome or atypical Rett syndrome. Here we report generation of the Cdkl5 knockout mouse and show that CDKL5 controls postsynaptic localization of GluN2B-containing N-methyl-d-aspartate (NMDA) receptors in the hippocampus and regulates seizure susceptibility. Cdkl5 -/Y mice showed normal sensitivity to kainic acid; however, they displayed significant hyperexcitability to NMDA. In concordance with this result, electrophysiological analysis in the hippocampal CA1 region disclosed an increased ratio of NMDA/α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated excitatory postsynaptic currents (EPSCs) and a significantly larger decay time constant of NMDA receptor-mediated EPSCs (NMDA-EPSCs) as well as a stronger inhibition of the NMDA-EPSCs by the GluN2B-selective antagonist ifenprodil in Cdkl5 -/Y mice. Subcellular fractionation of the hippocampus from Cdkl5 -/Y mice revealed a significant increase of GluN2B and SAP102 in the PSD (postsynaptic density)-1T fraction, without changes in the S1 (post-nuclear) fraction or mRNA transcripts, indicating an intracellular distribution shift of these proteins to the PSD. Immunoelectron microscopic analysis of the hippocampal CA1 region further confirmed postsynaptic overaccumulation of GluN2B and SAP102 in Cdkl5 -/Y mice. Furthermore, ifenprodil abrogated the NMDA-induced hyperexcitability in Cdkl5 -/Y mice, suggesting that upregulation of GluN2B accounts for the enhanced seizure susceptibility. These data indicate that CDKL5 plays an important role in controlling postsynaptic localization of the GluN2B-SAP102 complex in the hippocampus and thereby regulates seizure susceptibility, and that aberrant NMDA receptor-mediated synaptic transmission underlies the pathological mechanisms of the CDKL5 loss-of-function.

Keywords: CDKL5; Developmental disorders; Epilepsy; Excitatory synapses; GluN2B; Hippocampus; Ifenprodil; NMDA receptors; SAP102.

MeSH terms

Animals
Disease Models, Animal
Disease Susceptibility / metabolism
Excitatory Amino Acid Antagonists / pharmacology
Guanylate Kinases / metabolism
Hippocampus / drug effects
Hippocampus / metabolism*
Hippocampus / pathology
Kainic Acid
Membrane Proteins / metabolism
Mice, Inbred C57BL
Mice, Knockout
N-Methylaspartate
Piperidines / pharmacology
Post-Synaptic Density / drug effects
Post-Synaptic Density / metabolism*
Post-Synaptic Density / pathology
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism*
RNA, Messenger / metabolism
Receptors, N-Methyl-D-Aspartate / antagonists & inhibitors
Receptors, N-Methyl-D-Aspartate / metabolism*
Seizures / metabolism*
Seizures / pathology
Tissue Culture Techniques

Substances

Excitatory Amino Acid Antagonists
Membrane Proteins
NR2B NMDA receptor
Piperidines
RNA, Messenger
Receptors, N-Methyl-D-Aspartate
N-Methylaspartate
Protein Serine-Threonine Kinases
CDKL5 protein, mouse
Dlgh3 protein, mouse
Guanylate Kinases
ifenprodil
Kainic Acid