Choline oxidase chemiluminescent assay, after removal of eserine from medium, of acetylcholine released in vitro from brain slices.
A chemiluminescent method has been used recently for the determination of acetylcholine with limitations such as the presence of a cholinesterase inhibitor in the incubation medium, which is indispensable for the study of acetylcholine release by various agents. A modified procedure is presented in which the cholinesterase inhibitor eserine (physostigmine) is extracted from the medium. The results showed complete recovery when labelled acetylcholine was used. This modified procedure was used to determine the release of acetylcholine evoked by tityustoxin and ouabain. The results were comparable to those obtained by bioassay using a strip of guinea pig ileum.
Departamento de Bioquímica e Immunologia, Instituto de Ciências Biológicas UFMG, Belo Horizonte - Minas Gerais, Brazil.
M A
MA
Prado
T
T
Moraes-Santos
R N
RN
Freitas
M A
MA
Silva
M V
MV
Gomez
D000109:Acetylcholine / Q000032:analysis*; D000429:Alcohol Oxidoreductases / Q000032:analysis*; D000818:Animals; D001921:Brain / Q000201:enzymology*; D001923:Brain Chemistry; D010830:Physostigmine / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains
D010830:Physostigmine; D000429:Alcohol Oxidoreductases; C019888:choline oxidase; D000109:Acetylcholine