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Electrochemical Measurements of Optogenetically Stimulated Quantal Amine Release from Single Nerve Cell Varicosities in Drosophila Larvae
Author(s) -
Majdi Soodabeh,
Berglund E. Carina,
Dunevall Johan,
Oleinick Alexander I.,
Amatore Christian,
Krantz David E.,
Ewing Andrew G.
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201506743
Subject(s) - octopamine (neurotransmitter) , exocytosis , biogenic amine , drosophila melanogaster , biophysics , neuromuscular junction , synaptic vesicle , neurotransmitter , microelectrode , microbiology and biotechnology , chemistry , acetylcholine , ion channel , biology , biochemistry , vesicle , neuroscience , electrode , receptor , membrane , endocrinology , serotonin , gene
The nerve terminals found in the body wall of Drosophila melanogaster larvae are readily accessible to experimental manipulation. We used the light‐activated ion channel, channelrhodopsin‐2, which is expressed by genetic manipulation in Type II varicosities to study octopamine release in Drosophila. We report the development of a method to measure neurotransmitter release from exocytosis events at individual varicosities in the Drosophila larval system by amperometry. A microelectrode was placed in a region of the muscle containing a varicosity and held at a potential sufficient to oxidize octopamine and the terminal stimulated by blue light. Optical stimulation of Type II boutons evokes exocytosis of octopamine, which is detected through oxidization at the electrode surface. We observe 22700±4200 molecules of octopamine released per vesicle. This system provides a genetically accessible platform to study the regulation of amine release at an intact synapse.