Exclusive photorelease of signalling lipids at the plasma membrane

André Nadler; Dmytro A. Yushchenko; Rainer Müller; Frank Stein; Suihan Feng; Christophe Mulle; Mario Carta; Carsten Schultz

doi:10.1038/ncomms10056

Exclusive photorelease of signalling lipids at the plasma membrane

André Nadler, Dmytro A. Yushchenko, Rainer Müller, Frank Stein, Suihan Feng, Christophe Mulle, Mario Carta, Carsten Schultz

Research output: Contribution to journal › Article › peer-review

61 Scopus citations

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.

Original language	English (US)
Article number	10056
Journal	Nature communications
Volume	6
DOIs	https://doi.org/10.1038/ncomms10056
State	Published - Dec 21 2015
Externally published	Yes

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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title = "Exclusive photorelease of signalling lipids at the plasma membrane",

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.",

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note = "Funding Information: We thank the Advanced Light Microscopy Facility (ALMF) of European Molecular Biology Laboratory (EMBL) for expert support in microscopy. We thank the Miyazaki laboratory for kindly providing MIN6 cells and sharing information with regard to MIN6 cell culture. We thank S{\'e}bastien Marais at the Bordeaux Imaging Center, part of the FranceBioImaging national infrastructure, for support in two photon microscopy. This work was supported by the DFG (TRR83). D.A.Y. is a fellow of the EMBL Interdisciplinary Postdoc Programme (EIPOD).",

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AU - Müller, Rainer

AU - Stein, Frank

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AU - Mulle, Christophe

AU - Carta, Mario

AU - Schultz, Carsten

N1 - Funding Information: We thank the Advanced Light Microscopy Facility (ALMF) of European Molecular Biology Laboratory (EMBL) for expert support in microscopy. We thank the Miyazaki laboratory for kindly providing MIN6 cells and sharing information with regard to MIN6 cell culture. We thank Sébastien Marais at the Bordeaux Imaging Center, part of the FranceBioImaging national infrastructure, for support in two photon microscopy. This work was supported by the DFG (TRR83). D.A.Y. is a fellow of the EMBL Interdisciplinary Postdoc Programme (EIPOD).

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N2 - 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.

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Exclusive photorelease of signalling lipids at the plasma membrane

Abstract

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