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Inhibition of Calcineurin A by FK506 Suppresses Seizures and Reduces the Expression of GluN2B in Membrane Fraction

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Abstract

FK506, a calcineurin inhibitor, shows neuroprotective effects and has been associated with neurodegenerative diseases. Calcineurin A (CaNA), a catalytic subunit of calcineurin, mediates the dephosphorylation of various proteins. N-methyl-d-aspartate receptor (GluN) is closely related to epileptogenesis, and various phosphorylation sites of GluN2B, a regulatory subunit of the GluN complex, have different functions. Thus, we hypothesized that one of the potential anti-epileptic mechanisms of FK506 is mediated by its ability to promote the phosphorylation of GluN2B and reduce the expression of GluN2B in membrane fraction by down-regulating CaNA. CaNA expression was increased in the cortex of patients with temporal lobe epilepsy and pentylenetetrazol (PTZ)-induced epileptic models. CaNA was shown to be expressed in neurons using immunofluorescence staining. According to our behavioral observations, epileptic rats exhibited less severe seizures and were less sensitive to PTZ after a systemic injection of FK506. The levels of phosphorylated GluN2B were decreased in epileptic rats but increased after the FK506 treatment. Moreover, there was no difference in the total GluN2B levels before and after FK506 treatment. However, the expression of GluN2B in membrane fraction was suppressed after FK506 treatment. Based on these results, FK506 may reduce the severity and frequency of seizures by reducing the expression of GluN2B in membrane fraction.

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Abbreviations

CaN:

Calcineurin

CaNA:

Calcineurin A

GluN:

N-methyl-d-aspartate receptor

TLE:

Temporal lobe epilepsy.

PTZ:

Pentylenetetrazol

SRS:

Spontaneous recurrent seizures

GluRs:

Glutamate receptors

CaM:

Calmodulin

CREB:

cAMP response element-binding protein

NFAT:

Nuclear factor of activated T cells

BAD:

Bcl2-associated death promoter

EPSCs:

Excitatory postsynaptic currents

CsA:

Cyclosporin A

BDNF:

Brain-derived neurotrophic factor

GluA:

α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid

KA:

Kainic acid

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Acknowledgements

This work was supported by the Foundation of Chongqing Health Bureau (2013-1-005). We sincerely thank the support of Chongqing Key Laboratory of Neurology, which provided the brain tissue samples. We also thank the patients and families for the donation of brains.

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  1. Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, No.1, Youyi Road, Chongqing, 400010, China

    Yuetao Wen, Pengfei Fu, Kunlun Wu, Kaichuang Si, Yanfeng Xie, Wei Dan, Yan Zhan & Quanhong Shi

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  1. Yuetao Wen
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Correspondence to Quanhong Shi.

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All procedures performed in studies involving human participants were in accordance with the ethical standareds of the Chongqing Medical University and with the 1964 Helsinki declaration and its later amendments or comparable ethical standareds.

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Wen, Y., Fu, P., Wu, K. et al. Inhibition of Calcineurin A by FK506 Suppresses Seizures and Reduces the Expression of GluN2B in Membrane Fraction. Neurochem Res 42, 2154–2166 (2017). https://doi.org/10.1007/s11064-017-2221-0

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