Release of acetylcholine from murine embryonic stem cells: effect of nicotinic and muscarinic receptors and blockade of organic cation transporter

Life Sciences
I WesslerC J Kirkpatrick

Abstract

The non-neuronal cholinergic system is widely expressed in nature. The present experiments were performed to characterize the non-neuronal cholinergic system in murine embryonic stem cells (CGR8 cell line). CGR8 cells were cultured in gelatinized flasks with Glasgow's buffered minimal essential medium (Gibco, Germany). Acetylcholine was measured by HPLC combined with bioreactor and electrochemical detection. CGR8 cells contained 1.08±0.12 pmol acetylcholine/10(6) cells (n=7) which was reduced to 0.50±0.06 pmol/10(6) cells (n=6; p<0.05) in the presence (4h) of 30μM bromoacetylcholine to block choline acetyltransferase. A time-dependent release of acetylcholine into the incubation medium was demonstrated, when cholinesterase activity was blocked by 10 μM physostigmine, with 97±13, 180±15 and 216±14 pmol being released from 65×10(6) cells after incubation periods of 2, 4 and 6h, respectively. The cumulative release corresponds to a fractional release rate of 2%/min. Blockade of nicotine or muscarine receptors did not significantly modulate the release of acetylcholine which was substantially reduced by 300 μM quinine (inhibitor of organic cation transporters). This inhibition showed considerable fading over the incubation period, ...Continue Reading

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Citations

Feb 7, 2014·Respiratory Research·Ahmi Ben-YehudahDiane L Carlisle
Feb 14, 2014·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·Zu-Peng XuYong-Yao Cui
Nov 26, 2015·Expert Opinion on Drug Metabolism & Toxicology·Ágota ApátiBalázs Sarkadi
May 15, 2013·Neuroscience Letters·Ignaz WesslerCharles James Kirkpatrick
May 2, 2015·International Immunopharmacology·Ignaz WesslerCharles James Kirkpatrick
Aug 27, 2013·Experimental Physiology·Anastasia A BorodinovaGalina S Sukhova
Jan 11, 2017·Journal of Neurochemistry·Ignaz Karl Wessler, Charles James Kirkpatrick

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