Quantitative measurement of transmitters in individual vesicles in the cytoplasm of single cells with nanotip electrodes.
The quantification of vesicular transmitter content is important for studying the mechanisms of neurotransmission and malfunction in disease, and yet it is incredibly difficult to measure the tiny amounts of neurotransmitters in the attoliter volume of a single vesicle, especially in the cell environment. We introduce a novel method, intracellular vesicle electrochemical cytometry. A nanotip conical carbon-fiber microelectrode was used to electrochemically measure the total content of electroactive neurotransmitters in individual nanoscale vesicles in single PC12 cells as these vesicles lysed on the electrode inside the living cell. The results demonstrate that only a fraction of the quantal neurotransmitter content is released during exocytosis. These data support the intriguing hypothesis that the vesicle does not open all the way during the normal exocytosis process, thus resulting in incomplete expulsion of the vesicular contents.
Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemivägen 10, 41296 Gothenburg (Sweden).|Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 41296 Gothenburg (Sweden).
Xianchan
X
Li
Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemivägen 10, 41296 Gothenburg (Sweden).
Soodabeh
S
Majdi
Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemivägen 10, 41296 Gothenburg (Sweden).
Johan
J
Dunevall
Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemivägen 10, 41296 Gothenburg (Sweden).
Hoda
H
Fathali
Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Kemivägen 10, 41296 Gothenburg (Sweden). andrew.ewing@chem.gu.se.|Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 41296 Gothenburg (Sweden). andrew.ewing@chem.gu.se.
Andrew G
AG
Ewing
D000818:Animals; D002395:Catecholamines / Q000032:analysis*; D055664:Electrochemical Techniques / Q000295:instrumentation*; D004867:Equipment Design; D005089:Exocytosis; D008839:Microelectrodes; D018377:Neurotransmitter Agents / Q000032:analysis*; D016716:PC12 Cells; D051381:Rats; D059010:Single-Cell Analysis / Q000295:instrumentation*
D002395:Catecholamines; D018377:Neurotransmitter Agents
catecholamines; electrochemistry; exocytosis; nanoelectrodes; vesicles