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Reduced CaM Kinase II and CaM Kinase IV Activities Underlie Cognitive Deficits in NCKX2 Heterozygous Mice

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Abstract

Among five members of the K+-dependent Na+/Ca2+ exchanger (NCKX) family (NCKX1–5), only NCKX2 is highly expressed in mouse brain. NCKX2 in plasma membranes mediates cytosolic calcium excretion through electrogenic exchange of 4 Na+ for 1 Ca2+ and 1 K+. Here, we observed significantly decreased levels of NCKX2 protein and mRNA in the CA1 region of APP23 mice, a model of Alzheimer’s disease. We also found that, like APP23 mice, heterozygous NCKX2-mutant mice exhibit mildly impaired hippocampal LTP and memory acquisition, the latter based on novel object recognition and passive avoidance tasks. When we addressed underlying mechanisms, we found that both CaMKII autophosphorylation and CaMKIV phosphorylation significantly decreased in CA1 regions of NCKX2+/− relative to control mice. Likewise, phosphorylation of GluA1 (Ser-831) and CREB (Ser-133), respective downstream targets of CaMKII and CaMKIV, also significantly decreased in the CA1 region. BDNF protein and mRNA levels significantly decreased in CA1 of NCKX2+/− relative to control mice. Finally, CaN activity increased in CA1 of NCKX2+/− mice. Our findings suggest that like APP23 mice, NCKX2+/− mice may exhibit impaired learning and hippocampal LTP due to decreased CaM kinase II and CaM kinase IV activities.

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Abbreviations

AD:

Alzheimer’s disease

AMPAR:

α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor

BDNF:

Brain-derived neurotrophic factor

CaMKII:

Calcium/calmodulin-dependent protein kinase II

CaMKIV:

Calcium/calmodulin-dependent protein kinase IV

CaN:

Calcineurin

CREB:

cAMP-responsive element binding protein

DG:

Dentate gyrus

ERK:

Extracellular signal-regulated kinase

fEPSPs:

Field excitatory post-synaptic potentials

GFP:

Green fluorescent protein

HFS:

High-frequency stimulation

LTP:

Long-term potentiation

NCKXs:

K+-dependent Na+/Ca2+ exchangers

NCXs:

Na+/Ca2+ exchangers

PP1:

Protein phosphatase 1

WT:

Wild-type

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Acknowledgements

This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology, and the Ministry of Health and Welfare of Japan (KAKENHI 22390109 to K.F.; 20790398 to S.M.; 23590319 to T.I.; 25460350 to S.K.), the Uehara Memorial Foundation (K.F.) and the Smoking Research Foundation (S.M.). We also thank Novartis Pharma for providing APP23 mice.

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Authors and Affiliations

  1. Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aramaki-Aoba, Aoba-ku, Sendai, Miyagi, 980-8578, Japan

    Shigeki Moriguchi, Yasushi Yabuki, Ryo Inagaki, Hisanao Izumi, Yuzuru Sasaki & Kohji Fukunaga

  2. Department of Pharmacology, Faculty of Medicine, Fukuoka University, 7-45-1, Nanakuma, Jonan-ku, Fukuoka, Fukuoka, 814-0180, Japan

    Satomi Kita, Hideaki Tagashira & Takahiro Iwamoto

  3. Department of Pharmacology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima, Japan

    Satomi Kita

  4. Department of Physiology II, Nara Medical University, Nara, Japan

    Kyoji Horie

  5. Department of Social and Environmental Medicine, Graduate School of Medicine, Osaka University, Osaka, Japan

    Junji Takeda

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  1. Shigeki Moriguchi
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  2. Satomi Kita
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  3. Yasushi Yabuki
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Contributions

S.M., Y.Y., R.I., H.I., Y.S., K.S., H.T., and T.I. performed experiments. S.K., K.H., J.T., and T.I. provided NCKX2 antibody and NCKX2 knockout mice, and critical advice. S.M. and K.F. wrote the manuscript and designed the study.

Corresponding authors

Correspondence to Takahiro Iwamoto or Kohji Fukunaga.

Ethics declarations

All animal protocols were approved by the Committee on Animal Experiments at Tohoku University.

Conflict of Interest

The authors declare that they have no competing interests.

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Moriguchi, S., Kita, S., Yabuki, Y. et al. Reduced CaM Kinase II and CaM Kinase IV Activities Underlie Cognitive Deficits in NCKX2 Heterozygous Mice. Mol Neurobiol 55, 3889–3900 (2018). https://doi.org/10.1007/s12035-017-0596-1

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