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Proteomic analysis of mouse choroid plexus cell line ECPC-4 treated with lipid A

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Abstract

Objectives

Choroid plexus (CP) epithelial cells have multiple functions in the cerebral ventricles, including cerebrospinal fluid (CSF) production and forming part of the blood–CSF barrier. They are also responsible for producing inflammatory mediators involved in meningitis. The present study aimed to elucidate the functions of the CP epithelial cells during CNS inflammation.

Materials and methods

We analyzed the proteome and phosphoproteome in lipid A-treated ECPC-4 mouse CP cells by gel electrophoresis and mass spectrometry.

Results

Levels of 10 proteins and seven phosphoproteins were significantly altered by lipid A in time-dependent manners, including V-type proton ATPase subunit B (ATP6V), protein 40 kD, elongation factor-1δ, coatomer subunit ε (COPE), vimentin (isoform CRA a), purine nucleoside phosphorylase, eukaryotic initiation factor-4F splicing variant, put. β-actin, peroxiredoxin-6 isoform 1, and immunoglobulin heavy chain variable region. These proteins could be classified as having cytoskeleton/intermediate filament, protein-folding, signal-transduction, cell-growth, metabolism, and redox-regulation functions. The identified phosphoproteins were HSP 84, γ-actin, HSP 70 cognate, vimentin, tubulin β-4B chain, protein disulfide-isomerase A6 precursor, and heterogenous nuclear ribonucleoprotein, which could be classified as having cytoskeleton/intermediate filament, protein-folding, and metabolism functions.

Conclusions

These results indicate that lipid A can change the levels of proteins and phosphoproteins in ECPC-4 cells, suggesting that the identified proteins and phosphoproteins may play important roles in inflammation of the CP.

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Acknowledgments

This work was supported by grants from the Smoking Research Foundation, Kobe Gakuin University for Collaborative Research A, and Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (Nos. 18023033, 21500352, 21390271, 20023027).

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Authors and Affiliations

  1. Laboratory of Molecular Cellular Biology, School of Pharmaceutical Sciences, Kobe Gakuin University, 1-1-3 Minatojima, Chuo-ku, Kobe, 650-8586, Japan

    Masaoki Takano, Mieko Otani, Takahiro Kaji & Keiji Sano

  2. Laboratory of Physiology, School of Pharmaceutical Sciences, Kobe Gakuin University, 1-1-3 Minatojima, Chuo-ku, Kobe, 650-8586, Japan

    Michiko Hamada-Kanazawa Ph. D.

  3. Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences, Himeji Dokkyo University, 7-2-1 Kamiohno, Himeji, 670-8524, Japan

    Shogo Matsuyama

Authors
  1. Masaoki Takano
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  2. Mieko Otani
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  3. Takahiro Kaji
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  4. Keiji Sano
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  5. Michiko Hamada-Kanazawa Ph. D.
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  6. Shogo Matsuyama
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Correspondence to Masaoki Takano.

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Responsible Editor: John Di Battista.

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Takano, M., Otani, M., Kaji, T. et al. Proteomic analysis of mouse choroid plexus cell line ECPC-4 treated with lipid A. Inflamm. Res. 65, 295–302 (2016). https://doi.org/10.1007/s00011-016-0916-3

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  • DOI: https://doi.org/10.1007/s00011-016-0916-3

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