Get access
Advertisement

Localization and Dynamics of Glucocorticoid Receptor at the Plasma Membrane of Activated Mast Cells

Authors

  • Emmanuel Oppong,

    1. Karlsruhe Institute of Technology, Institute of Toxicology and Genetics, Eggenstein-Leopoldshafen, Germany
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Per Niklas Hedde,

    1. Karlsruhe Institute of Technology, Institute of Applied Physics and Centre for Functional Nanostructures, Karlsruhe, Germany
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Sylwia Sekula-Neuner,

    1. Karlsruhe Institute of Technology, Institute of Nanotechnology (INT) & Karlsruhe Nano Micro Facility (KNMF), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany
    Search for more papers by this author
  • Linxiao Yang,

    1. Karlsruhe Institute of Technology, Institute of Applied Physics and Centre for Functional Nanostructures, Karlsruhe, Germany
    Search for more papers by this author
  • Falko Brinkmann,

    1. Karlsruhe Institute of Technology, Institute of Nanotechnology (INT) & Karlsruhe Nano Micro Facility (KNMF), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany
    2. Physical Institute and Center for Nanotechnology (CeNTech) University of Münster, Münster, Germany
    Search for more papers by this author
  • René M. Dörlich,

    1. Karlsruhe Institute of Technology, Institute of Applied Physics and Centre for Functional Nanostructures, Karlsruhe, Germany
    Search for more papers by this author
  • Michael Hirtz,

    1. Karlsruhe Institute of Technology, Institute of Nanotechnology (INT) & Karlsruhe Nano Micro Facility (KNMF), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany
    Search for more papers by this author
  • Harald Fuchs,

    1. Karlsruhe Institute of Technology, Institute of Nanotechnology (INT) & Karlsruhe Nano Micro Facility (KNMF), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany
    2. Physical Institute and Center for Nanotechnology (CeNTech) University of Münster, Münster, Germany
    Search for more papers by this author
  • Gerd Ulrich Nienhaus,

    1. Karlsruhe Institute of Technology, Institute of Toxicology and Genetics, Eggenstein-Leopoldshafen, Germany
    2. Karlsruhe Institute of Technology, Institute of Applied Physics and Centre for Functional Nanostructures, Karlsruhe, Germany
    3. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, USA
    Search for more papers by this author
  • Andrew C. B. Cato

    Corresponding author
    1. Karlsruhe Institute of Technology, Institute of Toxicology and Genetics, Eggenstein-Leopoldshafen, Germany
    Search for more papers by this author

Abstract

In addition to their actions in the cell nucleus, glucocorticoids exhibit rapid non-nuclear responses that are mechanistically not well understood. To explain these effects, the localization of a glucocorticoid receptor (GR) expressed in mast cells as a GFP fusion was analyzed after activation of the cells on allergenic lipid arrays. These arrays were produced on glass slides by dip-pen nanolithography (DPN) and total internal reflection (TIRF) microscopy was used to visualize the GR. A rapid glucocorticoid-independent and -dependent recruitment of the GR-GFP to the plasma cell membrane was observed following contact of the cells with the allergenic array. In addition, the mobility of the GR at the membrane was monitored by fluorescence recovery after photobleaching (FRAP) and shown to follow binding kinetics demonstrating interactions of the receptor with membrane-bound factors. Furthermore the recruitment of the GR to the cell membrane was shown to result in a glucocorticoid-mediated increase in Erk phosphorylation. This is evidenced by findings that destruction of the membrane composition of the mast cells by cholesterol depletion impairs the membrane localization of the GR and subsequent glucocorticoid-mediated enhancement of Erk phosphorylation. These results demonstrate a membrane localization and function of the GR in mast cell signaling.

Get access to the full text of this article

Ancillary