Exclusive photorelease of signalling lipids at the plasma membrane

André Nadler, Dmytro A. Yushchenko, Rainer Müller, Frank Stein, Suihan Feng, Christophe Mulle, Mario Carta and Carsten Schultz ()
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André Nadler: European Molecular Biology Laboratory, Cell Biology and Biophysics Unit
Dmytro A. Yushchenko: European Molecular Biology Laboratory, Cell Biology and Biophysics Unit
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

Nature Communications, 2015, vol. 6, issue 1, 1-10

Abstract: 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.

Date: 2015
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DOI: 10.1038/ncomms10056

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