Mechanistic Aspects of Vesicle Opening during Analysis with Vesicle Impact Electrochemical Cytometry
Artikel i vetenskaplig tidskrift, 2017

Vesicle impact electrochemical cytometry (VIEC) has been used to quantify the vesicular transmitter content in mammalian vesicles. In the present study, we studied the mechanism of VIEC by quantifying the catecholamine content in single vesicles isolated from pheochromocytoma (PC12) cells. These vesicles contain about one tenth of the catecholamine compared with adrenal chromaffin vesicles. The existence of a prespike foot for many events suggests the formation of an initial transiently stable pore at the beginning of vesicle rupture. Increasing the detection temperature from 6 to 30 °C increases the possibility of vesicle rupture on the electrode, implying that there is a temperature-dependent process that facilitates electroporation. Natively larger vesicles are shown to rupture earlier and more frequently than smaller ones in VIEC. Likewise, manipulating vesicle content and size with drugs leads to similar trends. These data support the hypothesis that electroporation is the primary force for pore opening in VIEC. We further hypothesize that a critical step for initiating vesicle opening by electroporation is diffusion of membrane proteins away from the membrane region of contact with the electrode to allow closer contact, increasing the lateral potential field and thus facilitating electroporation.

Författare

Li Xianchan

Göteborgs universitet

Johan Dunevall

Kemi och kemiteknik, Kemi och biokemi, Analytisk kemi

Lin Ren

Kemi och kemiteknik, Kemi och biokemi, Analytisk kemi

Andrew Ewing

Kemi och kemiteknik, Kemi och biokemi, Analytisk kemi

Analytical Chemistry

0003-2700 (ISSN) 1520-6882 (eISSN)

Vol. 89 9416-9423

Ämneskategorier

Analytisk kemi

DOI

10.1021/acs.analchem.7b02226