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Nano Secondary Ion Mass Spectrometry Imaging of Dopamine Distribution Across Nanometer Vesicles

Journal article
Authors Jelena Lovrić
Johan Dunevall
Anna Larsson
Lin Ren
S. Andersson
A. Meibom
Per Malmberg
M.E. Kurczy
Andrew G Ewing
Published in ACS Nano
Volume 11
Issue 4
Pages 3446-3455
ISSN 1936-0851
Publication year 2017
Published at Department of Chemistry and Molecular Biology
National Center for Imaging Mass Spectrometry
Pages 3446-3455
Language en
Links dx.doi.org/10.1021/acsnano.6b07233
Keywords electrochemistry; nanocompartments; nanoimaging; NanoSIMS; vesicle content
Subject categories Analytical Chemistry

Abstract

We report an approach to spatially resolve the content across nanometer neuroendocrine vesicles in nerve-like cells by correlating super high-resolution mass spectrometry imaging, NanoSIMS, with transmission electron microscopy (TEM). Furthermore, intracellular electrochemical cytometry at nanotip electrodes is used to count the number of molecules in individual vesicles to compare to imaged amounts in vesicles. Correlation between the NanoSIMS and TEM provides nanometer resolution of the inner structure of these organelles. Moreover, correlation with electrochemical methods provides a means to quantify and relate vesicle neurotransmitter content and release, which is used to explain the slow transfer of dopamine between vesicular compartments. These nanoanalytical tools reveal that dopamine loading/unloading between vesicular compartments, dense core and halo solution, is a kinetically limited process. The combination of NanoSIMS and TEM has been used to show the distribution profile of newly synthesized dopamine across individual vesicles. Our findings suggest that the vesicle inner morphology might regulate the neurotransmitter release event during open and closed exocytosis from dense core vesicles with hours of equilibrium needed to move significant amounts of catecholamine from the protein dense core despite its nanometer size.

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