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Journal Article

Exclusive photorelease of signalling lipids at the plasma membrane.

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Nadler,  André
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Citation

Nadler, A., Yushchenko, D. A., Müller, R., Stein, F., Feng, S., Mulle, C., et al. (2015). Exclusive photorelease of signalling lipids at the plasma membrane. Nature Communications, 6: 10056.


Cite as: https://hdl.handle.net/21.11116/0000-0001-042C-A
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.