Volume 210, 2018

Electrochemical quantification of transmitter concentration in single nanoscale vesicles isolated from PC12 cells

Abstract

We use an electrochemical platform, nanoparticle tracking analysis, and differential centrifugation of single catecholamine vesicles to study the properties of nanometer transmitter vesicles, including the number of molecules, size, and catecholamine concentration inside. Vesicle impact electrochemical cytometry (VIEC) was used to quantify the catecholamine content of single vesicles in different batches isolated from pheochromocytoma (PC12) cells with different ultracentrifugation speeds. We show that, vesicles containing less catecholamine are obtained at subsequent centrifugation steps with higher speed (force). Important to quantification, the cumulative content after subsequent centrifugation steps is equivalent to that of one-step centrifugation at the highest speed, 70 000g. Moreover, as we count molecules in the vesicles, we compared molecular numbers from VIEC, flow VIEC, and intracellular VIEC to corresponding vesicle size measured by nanoparticle tracking analysis to evaluate catecholamine concentration in vesicles. The data suggest that vesicular catecholamine concentration is relatively constant and independent of the vesicular size, indicating vesicular transmitter content as a main factor regulating the vesicle size.

Graphical abstract: Electrochemical quantification of transmitter concentration in single nanoscale vesicles isolated from PC12 cells

Associated articles

Supplementary files

Article information

Article type
Paper
Submitted
08 फरवरी 2018
Accepted
08 मार्च 2018
First published
08 मार्च 2018

Faraday Discuss., 2018,210, 353-364

Electrochemical quantification of transmitter concentration in single nanoscale vesicles isolated from PC12 cells

X. Li, J. Dunevall and A. G. Ewing, Faraday Discuss., 2018, 210, 353 DOI: 10.1039/C8FD00020D

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements