Monitoring of acetylcholine released from postnatal rat basal forebrain cholinergic neurons cultured on membrane filter by cell bed perfusion system and HPLC-ECD.
A method for the determination of release of endogenous neurotransmitter from cultured neurons has been developed. Basal forebrain cholinergic neurons from postnatal rats were cultured on polyethyleneimine-coated membrane filter with nerve growth factor. The membrane filter, on which the neurons lived and extended neurites, was packed in a cell bed chamber for a closed perfusion. The perfusate was fractionated and the content of acetylcholine released from cultured neurons was measured by high-performance liquid chromatography-electrochemical detection (HPLC-ECD). Acetylcholine release evoked by glutamate and high K+ was 1.5- and 3-fold greater than the spontaneous level. This method for the determination of the neurotransmitter released from cultured neurons is very advantageous for investigating regulation of transmitter release in central neurons.
Takei
N
N
Department of Neuroscience, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Tsukui
H
H
Kumakura
K
K
Hatanaka
H
H
D000109:Acetylcholine / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D002799:Cholinergic Fibers / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004027:Diencephalon / Q000166:cytology / Q000378:metabolism*; D005374:Filtration / Q000295:instrumentation; D009474:Neurons / Q000166:cytology; D051381:Rats; D013687:Telencephalon / Q000166:cytology / Q000378:metabolism*
D000109:Acetylcholine