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Homostachydrine is a Xenobiotic Substrate of OCTN1/SLC22A4 and Potentially Sensitizes Pentylenetetrazole-Induced Seizures in Mice

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Abstract

Understanding of the underlying mechanism of epilepsy is desired since some patients fail to control their seizures. The carnitine/organic cation transporter OCTN1/SLC22A4 is expressed in brain neurons and transports food-derived antioxidant ergothioneine (ERGO), l-carnitine, and spermine, all of which may be associated with epilepsy. This study aimed to clarify the possible association of this transporter with epileptic seizures. In both pentylenetetrazole (PTZ)-induced acute seizure and kindling models, ocnt1 gene knockout mice (octn1−/−) showed lower seizure scores compared with wild-type mice. Up-regulation of the epilepsy-related genes, c-fos and Arc, and the neurotrophic factor BDNF following PTZ administration was observed in the hippocampus of wild-type, but not octn1−/− mice. To find the OCTN1 substrate associated with the seizure, untargeted metabolomics analysis using liquid chromatography–quadrupole time-of-flight mass spectrometry was conducted on extracts from the hippocampus, frontal cortex, and plasma of both strains, leading to the identification of a plant alkaloid homostachydrine as a compound present in a lower concentration in octn1−/− mice. OCTN1-mediated uptake of deuterium-labeled homostachydrine was confirmed in OCTN1-transfected HEK293 cells, suggesting that this compound is a substrate of OCTN1. Homostachydrine administration increased PTZ-induced acute seizure scores and the expression of Arc in the hippocampus and that of Arc, Egr1, and BDNF in the frontal cortex. Conversely, administration of the OCTN1 substrate/inhibitor ERGO inhibited PTZ-induced kindling and reduced the plasma homostachydrine concentration. Thus, these results suggest that OCTN1 is at least partially associated with PTZ-induced seizures, which is potentially deteriorated by treatment with homostachydrine, a newly identified food-derived OCTN1 substrate.

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Data Availability

The datasets used or analyzed during this study are available from the corresponding author upon reasonable request.

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Funding

Grants-in-Aid for Scientific Research to Noritaka Nakamichi (No. 19K07126) and Yukio Kato (No. 15H04664) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and grant by the Hoansha Foundation (Osaka, Japan) to Yukio Kato.

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Authors and Affiliations

  1. Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, 920-1192, Japan

    Misa Nishiyama, Noritaka Nakamichi, Tomoyuki Yoshimura, Yusuke Masuo, Tomoe Komori, Takahiro Ishimoto, Jun-ichi Matsuo & Yukio Kato

  2. Faculty of Pharmacy, Takasaki University of Health and Welfare, 60 Nakaorui-machi, Takasaki, Gunma, 370-0033, Japan

    Noritaka Nakamichi

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  1. Misa Nishiyama
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  2. Noritaka Nakamichi
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Contributions

Conceptualization: MN, NN, YK. Methodology: NN, TY, YM, YK. Analysis and investigation: MN, TY, TK, YM. Writing—original draft preparation: MN, TK, TY. Writing, review, and editing: NN, TI, YK. Funding acquisition: NN, YK. Resources: YK. Supervision: NN, JM, YK.

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Correspondence to Noritaka Nakamichi.

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The authors declare that they have no competing interest.

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Experiments were performed according to the Guidelines for the Care and Use of Laboratory Animals at Kanazawa University. All protocols were approved by the Institutional Animal Care and Use Committee of Kanazawa University.

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Nishiyama, M., Nakamichi, N., Yoshimura, T. et al. Homostachydrine is a Xenobiotic Substrate of OCTN1/SLC22A4 and Potentially Sensitizes Pentylenetetrazole-Induced Seizures in Mice. Neurochem Res 45, 2664–2678 (2020). https://doi.org/10.1007/s11064-020-03118-8

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