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Exclusive photorelease of signalling lipids at the plasma membrane

Author

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  • André Nadler

    (European Molecular Biology Laboratory, Cell Biology and Biophysics Unit
    Max Planck Institute of Molecular Cell Biology and Genetics)

  • Dmytro A. Yushchenko

    (European Molecular Biology Laboratory, Cell Biology and Biophysics Unit
    Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic)

  • Rainer Müller

    (European Molecular Biology Laboratory, Cell Biology and Biophysics Unit)

  • Frank Stein

    (European Molecular Biology Laboratory, Cell Biology and Biophysics Unit)

  • Suihan Feng

    (European Molecular Biology Laboratory, Cell Biology and Biophysics Unit)

  • Christophe Mulle

    (Institut Interdisciplinaire de Neurosciences, CNRS UMR 5297 Université Bordeaux 2)

  • Mario Carta

    (Institut Interdisciplinaire de Neurosciences, CNRS UMR 5297 Université Bordeaux 2)

  • Carsten Schultz

    (European Molecular Biology Laboratory, Cell Biology and Biophysics Unit)

Abstract

Photoactivation of caged biomolecules has become a powerful approach to study cellular signalling events. Here we report a method for anchoring and uncaging biomolecules exclusively at the outer leaflet of the plasma membrane by employing a photocleavable, sulfonated coumarin derivative. The novel caging group allows quantifying the reaction progress and efficiency of uncaging reactions in a live-cell microscopy setup, thereby greatly improving the control of uncaging experiments. We synthesized arachidonic acid derivatives bearing the new negatively charged or a neutral, membrane-permeant coumarin caging group to locally induce signalling either at the plasma membrane or on internal membranes in β-cells and brain slices derived from C57B1/6 mice. Uncaging at the plasma membrane triggers a strong enhancement of calcium oscillations in β-cells and a pronounced potentiation of synaptic transmission while uncaging inside cells blocks calcium oscillations in β-cells and causes a more transient effect on neuronal transmission, respectively. The precise subcellular site of arachidonic acid release is therefore crucial for signalling outcome in two independent systems.

Suggested Citation

  • André Nadler & Dmytro A. Yushchenko & Rainer Müller & Frank Stein & Suihan Feng & Christophe Mulle & Mario Carta & Carsten Schultz, 2015. "Exclusive photorelease of signalling lipids at the plasma membrane," Nature Communications, Nature, vol. 6(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms10056
    DOI: 10.1038/ncomms10056
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