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A Mechanism Enhancing Macromolecule Transport Through Paracellular Spaces Induced by Poly-L-Arginine: Poly-L-Arginine Induces the Internalization of Tight Junction Proteins via Clathrin-Mediated Endocytosis

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ABSTRACT

Purpose

Poly-L-arginine (PLA) enhances the paracellular permeability of the Caco-2 cell monolayer to hydrophilic macromolecules by disappearance of tight junction (TJ) proteins from cell–cell junctions. However, the mechanism of the disappearance of TJ proteins in response to PLA has been unclear. In this study, we investigated the mechanism of disappearance of TJ proteins from cell–cell junctions after the application of PLA to Caco-2 cell monolayers.

Methods

The membrane conductance (Gt), FITC-dextran (FD-4) permeability, and localization of TJ proteins were examined after the treatment of Caco-2 cell monolayers with PLA in the presence of various endocytosis inhibitors. In addition, the localization of endosome marker proteins was also observed.

Results

Clathrin-mediated endocytosis inhibitors suppressed the increase in Gt and Papp of FD-4 induced by PLA, and also significantly suppressed the disappearance of TJ proteins induced by PLA. Furthermore, occludin, one of the TJ proteins, colocalized with early endosome and recycling endosomes after the internalization of occludin induced by PLA, and then was recycled to the cell–cell junctions.

Conclusion

PLA induced the transient internalization of TJ proteins in cell–cell junctions via clathrin-mediated endocytosis, subsequently increasing the permeability of the Caco-2 cell monolayer to FD-4 via a paracellular route.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama, 350-0295, Japan

    Tsutomu Yamaki, Yusuke Kamiya, Kazuo Ohtake, Masaki Uchida, Toshinobu Seki, Hideo Ueda, Jun Kobayashi, Yasunori Morimoto & Hideshi Natsume

  2. Research Institute of TTS Technology, Josai University, 1-1 Keyakidai, Sakado, Saitama, 350-0295, Japan

    Toshinobu Seki, Yasunori Morimoto & Hideshi Natsume

Authors
  1. Tsutomu Yamaki
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  2. Yusuke Kamiya
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  3. Kazuo Ohtake
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  4. Masaki Uchida
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  5. Toshinobu Seki
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  6. Hideo Ueda
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  7. Jun Kobayashi
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  8. Yasunori Morimoto
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  9. Hideshi Natsume
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Correspondence to Hideshi Natsume.

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Yamaki, T., Kamiya, Y., Ohtake, K. et al. A Mechanism Enhancing Macromolecule Transport Through Paracellular Spaces Induced by Poly-L-Arginine: Poly-L-Arginine Induces the Internalization of Tight Junction Proteins via Clathrin-Mediated Endocytosis. Pharm Res 31, 2287–2296 (2014). https://doi.org/10.1007/s11095-014-1324-4

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  • DOI: https://doi.org/10.1007/s11095-014-1324-4

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