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Turnover Rate of Brain Acetylcholine Using HPLC Separation of the Transmitter
Author(s) -
Bertrand N.,
Bralet J.,
Beley A.
Publication year - 1990
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1990.tb08816.x
Subject(s) - chemistry , acetylcholine , chromatography , oxotremorine , high performance liquid chromatography , choline , choline oxidase , cholinesterase , muscarinic acetylcholine receptor , biochemistry , enzyme , acetylcholinesterase , medicine , receptor
A simple, reliable method was developed for measuring brain acetylcholine (ACh) turnover using HPLC methodology. Mice were injected intravenously with [ 3 H]choline ([ 3 H]Ch), and the turnover rate of ACh was calculated from the formation of [ 3 H]ACh. Ch and ACh were separated from phosphorylcholine and from other radioactive compounds using tetraphenylboron extraction and counterion/reverse‐phase chromatography. Endogenous Ch and ACh were quantified electrochemically through hydrogen peroxide production in a postcolumn reactor containing covalently bonded ACh esterase and Ch oxidase. Labeled Ch and ACh were quantified in the same sample by collecting the chromatographic fractions for radioactive content determinations. The method is rapid, well adapted to large series, and highly reproducible, with recoveries of 72.1% for Ch and 79.3% for ACh. The turnover value in mouse cerebral hemispheres was 16.02 nmol g ‐1 min ‐1 and decreased to 9.94 nmol g ‐1 min ‐1 in mice treated with oxotremorine.