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PMID: 29068313 Published · epublish English Journal Article Research Support, Non-U.S. Gov't Research Support, N.I.H., Extramural

Phosphatidylinositol 4,5-bisphosphate optical uncaging potentiates exocytosis.

eLife ·Vol. 6 ·2017-00-25

Walter AM, Müller R, Tawfik B, Wierda KD, Pinheiro PS, Nadler A, McCarthy AW, Ziomkiewicz I, Kruse M, Reither G, Rettig J, Lehmann M, Haucke V, Hille B, Schultz C, Sørensen JB

Abstract

Phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] is essential for exocytosis. Classical ways of manipulating PI(4,5)P2 levels are slower than its metabolism, making it difficult to distinguish effects of PI(4,5)P2 from those of its metabolites. We developed a membrane-permeant, photoactivatable PI(4,5)P2, which is loaded into cells in an inactive form and activated by light, allowing sub-second increases in PI(4,5)P2 levels. By combining this compound with electrophysiological measurements in mouse adrenal chromaffin cells, we show that PI(4,5)P2 uncaging potentiates exocytosis and identify synaptotagmin-1 (the Ca2+ sensor for exocytosis) and Munc13-2 (a vesicle priming protein) as the relevant effector proteins. PI(4,5)P2 activation of exocytosis did not depend on the PI(4,5)P2-binding CAPS-proteins, suggesting that PI(4,5)P2 uncaging may bypass CAPS-function. Finally, PI(4,5)P2 uncaging triggered the rapid fusion of a subset of readily-releasable vesicles, revealing a rapid role of PI(4,5)P2 in fusion triggering. Thus, optical uncaging of signaling lipids can uncover their rapid effects on cellular processes and identify lipid effectors.

Keywords
Munc13 adrenal chromaffin cell cell biology exocytosis mouse neuroscience optical uncaging phosphatidylinositols synaptotagmin
MeSH Terms
Animals Carrier Proteins/metabolism Cell Line Chromaffin Cells/metabolism Cytological Techniques/methods Exocytosis Intracellular Signaling Peptides and Proteins/metabolism Membrane Proteins/metabolism Mice Nerve Tissue Proteins/metabolism Phosphatidylinositol 4,5-Diphosphate/metabolism Synaptotagmin I/metabolism
Chemicals
Carrier Proteins Cybr protein, mouse Intracellular Signaling Peptides and Proteins Membrane Proteins Nerve Tissue Proteins Phosphatidylinositol 4,5-Diphosphate Synaptotagmin I Syt1 protein, mouse Unc13b protein, mouse
Authors & Affiliations
16 authors, click to expand affiliations / ORCID
Walter Alexander M ORCID
Neurosecretion group, Center for Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. | Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany.
Müller Rainer ORCID
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Tawfik Bassam ORCID
Neurosecretion group, Center for Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Wierda Keimpe Db ORCID
Neurosecretion group, Center for Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Pinheiro Paulo S
Neurosecretion group, Center for Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Nadler André
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany. | Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
McCarthy Anthony W ORCID
Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany.
Ziomkiewicz Iwona
Neurosecretion group, Center for Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. | Discovery Sciences, AstraZeneca, Cambridge, United Kingdom.
Kruse Martin
Department of Physiology and Biophysics, School of Medicine, University of Washington, Seattle, United States.
Reither Gregor
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Rettig Jens
Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine, Saarland University, Homburg, Germany.
Lehmann Martin
Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany.
Haucke Volker
Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Berlin, Germany.
Hille Bertil
Department of Physiology and Biophysics, School of Medicine, University of Washington, Seattle, United States.
Schultz Carsten
Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Sørensen Jakob Balslev ORCID
Neurosecretion group, Center for Neuroscience, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
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Article Info
Journal
eLife
Abbr.
Elife
ISSN
2050-084X
DOI
10.7554/eLife.30203
Published
2017-00-25
Epub
2017-00-25
Language
English
Region
England
NLM ID
101579614
PMCID
PMC5711374
Subset
IM
Grants
NINDS NIH HHS · R37 NS008174 · United States
External Links
PubMed source DOI: 10.7554/eLife.30203 PMC Full Text View journal details
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