Localized Topological Changes of the Plasma Membrane Upon Exocytosis Visualized by Polarized TIRFM

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2010 Feb 10

PMID 20142424

Citations 70

Authors

Arun Anantharam

Bibiana Onoa

Robert H Edwards

Ronald W Holz

Daniel Axelrod

Affiliations

Soon will be listed here.

Abstract

Total internal reflection fluorescence microscopy (TIRFM) images the plasma membrane-cytosol interface and has allowed insights into the behavior of individual secretory granules before and during exocytosis. Much less is known about the dynamics of the other partner in exocytosis, the plasma membrane. In this study, we report the implementation of a TIRFM-based polarization technique to detect rapid submicrometer changes in plasma membrane topology as a result of exocytosis. A theoretical analysis of the technique is presented together with image simulations of predicted topologies of the postfusion granule membrane-plasma membrane complex. Experiments on diI-stained bovine adrenal chromaffin cells using polarized TIRFM demonstrate rapid and varied submicrometer changes in plasma membrane topology at sites of exocytosis that occur immediately upon fusion. We provide direct evidence for a persistent curvature in the exocytotic region that is altered by inhibition of dynamin guanosine triphosphatase activity and is temporally distinct from endocytosis measured by VMAT2-pHluorin.

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