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Neurochemical Research

, Volume 43, Issue 1, pp 116–128 | Cite as

Carnitine/Organic Cation Transporter OCTN1 Negatively Regulates Activation in Murine Cultured Microglial Cells

  • Takahiro Ishimoto
  • Noritaka NakamichiEmail author
  • Hikari Nishijima
  • Yusuke Masuo
  • Yukio Kato
Original Paper

Abstract

Brain immune cells, i.e., microglia, play an important role in the maintenance of brain homeostasis, whereas chronic overactivation of microglia is involved in the development of various neurodegenerative disorders. Therefore, the regulation of microglial activation may contribute to their treatment. The aim of the present study was to clarify the functional expression of carnitine/organic cation transporter OCTN1/SLC22A4, which recognizes the naturally occurring food-derived antioxidant ergothioneine (ERGO) as a substrate in vivo, in microglia and its role in regulation of microglial activation. Primary cultured microglia derived from wild-type mice (WT-microglia) and mouse microglial cell line BV2 exhibited time-dependent uptake of [3H]- or d9-labeled ERGO. The uptake was markedly decreased in cultured microglia from octn1 gene knockout mice (octn1−/−-microglia) and BV2 cells transfected with small interfering RNA targeting the mouse octn1 gene (siOCTN1). These results demonstrate that OCTN1 is functionally expressed in murine microglial cells. Exposure of WT-microglia to ERGO led to a significant decrease in cellular hypertrophy by LPS-stimulation with concomitant attenuation of intracellular reactive oxygen species (ROS), suggesting that OCTN1-mediated ERGO uptake may suppress cellular hypertrophy via the inhibition of ROS production with microglial activation. The expression of mRNA for interleukin-1β (IL-1β) after LPS-treatment was significantly increased in octn1−/−-microglia and siOCTN1-treated BV2 cells compared to the control cells. Meanwhile, treatment of ERGO minimally affected the induction of IL-1β mRNA by LPS-stimulation in cultured microglia and BV2 cells. Thus, OCTN1 negatively regulated the induction of inflammatory cytokine IL-1β, at least in part, via the transport of unidentified substrates other than ERGO in microglial cells.

Keywords

Microglia OCTN1 Ergothioneine Hypertrophy Interleukin-1β 

Abbreviations

DMEM

Dulbecco’s modified Eagle’s medium

ERGO

Ergothioneine

FBS

Fetal bovine serum

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

IL-1β

Interleukin-1β

LC-MS/MS

Liquid chromatography-mass spectrometry/mass spectrometry

LPS

Lipopolysaccharide

MRM

Multiple reaction monitoring

OCTN1

Carnitine/organic cation transporter 1

octn1−/−

octn1 gene knockout

octn1−/−-microglia

Microglia derived from octn1−/− mice

PBS

Phosphate-buffered saline

ROS

Reactive oxygen species

RT-PCR

Reverse transcription polymerase chain reaction

SLC

Solute carrier

siRNA

Small interfering RNA

siOCTN1

SiRNA targeting the mouse octn1gene

TNFα

Tumor necrosis factor-α

WT-microglia

Microglia derived from wild-type mice

Notes

Acknowledgements

This work was partially supported by Grants-in-Aid for Scientific Research to NN (No. 16K08266) and YK (No. 15H04664) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health SciencesKanazawa UniversityKanazawaJapan

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