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

Neurobiology of Disease (Oct 2017)

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

Affiliations

Kosuke Okuda: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Shizuka Kobayashi: Division of Neuronal Network, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
Masahiro Fukaya: Department of Anatomy, Kitasato University School of Medicine, Sagamihara 252-0374, Japan
Aya Watanabe: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Takuto Murakami: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Mai Hagiwara: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Tempei Sato: Department of Systems Biomedicine, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan
Hiroe Ueno: Department of Systems Biomedicine, National Research Institute for Child Health and Development, Tokyo 157-8535, Japan
Narumi Ogonuki: Bioresource Engineering Division, RIKEN BioResource Center, Tsukuba 305-0074, Japan
Sayaka Komano-Inoue: Department of Physiology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Hiroyuki Manabe: Department of Physiology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Masahiro Yamaguchi: Department of Physiology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Atsuo Ogura: Bioresource Engineering Division, RIKEN BioResource Center, Tsukuba 305-0074, Japan
Hiroshi Asahara: 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
Hiroyuki Sakagami: Department of Anatomy, Kitasato University School of Medicine, Sagamihara 252-0374, Japan
Masashi Mizuguchi: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan
Toshiya Manabe: Division of Neuronal Network, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
Teruyuki Tanaka: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan; Corresponding author at: Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Journal volume & issue: Vol. 106
pp. 158 – 170

Abstract

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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.

Published in Neurobiology of Disease

ISSN: 0969-9961 (Print); 1095-953X (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Medicine: Internal medicine: Neurosciences. Biological psychiatry. Neuropsychiatry
Website: https://www.journals.elsevier.com/neurobiology-of-disease

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