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Göteborgs universitets publikationer

Amperometric Detection of Single Vesicle Acetylcholine Release Events from an Artificial Cell

Författare och institution:
Jacqueline Keighron (Institutionen för kemi och kemiteknik, Analytisk kemi, Chalmers); Joakim Wigström (Institutionen för kemi och kemiteknik, Analytisk kemi, Chalmers); Michael E. Kurczy (Institutionen för kemi och kemiteknik, Analytisk kemi, Chalmers); Jenny Bergman (Institutionen för kemi och molekylärbiologi); Yuanmo Wang (Institutionen för kemi och kemiteknik, Analytisk kemi, Chalmers); Ann-Sofie Cans (Institutionen för kemi och kemiteknik, Analytisk kemi, Chalmers)
Publicerad i:
ACS Chemical Neuroscience, 6 ( 1 ) s. 181-188
ISSN:
1948-7193
Publikationstyp:
Artikel, refereegranskad vetenskaplig
Publiceringsår:
2015
Språk:
engelska
Fulltextlänk:
Sammanfattning (abstract):
Acetylcholine is a highly abundant nonelectroactive neurotransmitter in the mammalian central nervous system. Neurochemical release occurs on the millisecond time scale, requiring a fast, sensitive sensor such as an enzymatic amperometric electrode. Typically, the enzyme used for enzymatic electrochemical sensors is applied in excess to maximize signal. Here, in addition to sensitivity, we have also sought to maximize temporal resolution, by designing a sensor that is sensitive enough to work at near monolayer enzyme coverage. Reducing the enzyme layer thickness increases sensor temporal resolution by decreasing the distance and reducing the diffusion time for the enzyme product to travel to the sensor surface for detection. In this instance, the sensor consists of electrodeposited gold nanoparticle modified carbon fiber microelectrodes (CFMEs). Enzymes often are sensitive to curvature upon surface adsorption; thus, it was important to deposit discrete nanoparticles to maintain enzyme activity while depositing as much gold as possible to maximize enzyme coverage. To further enhance sensitivity, the enzymes acetylcholinesterase (AChE) and choline oxidase (ChO) were immobilized onto the gold nanoparticles at the previously determined optimal ratio (1:10 AChE/ChO) for most efficient sequential enzymatic activity. This optimization approach has enabled the rapid detection to temporally resolve single vesicle acetylcholine release from an artificial cell. The sensor described is a significant advancement in that it allows for the recording of acetylcholine release on the order of the time scale for neurochemical release in secretory cells.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
NATURVETENSKAP ->
Biologiska vetenskaper ->
Biokemi och molekylärbiologi
Nyckelord:
Biosensor, acetylcholine, amperometry, artificial cell, enzymes, exocytosis, gold nanoparticles
Postens nummer:
213331
Ingår i post nr:
Posten skapad:
2015-03-03 11:00
Posten ändrad:
2016-08-22 07:34

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