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首页> 外文期刊>Life sciences >Release of acetylcholine from murine embryonic stem cells: effect of nicotinic and muscarinic receptors and blockade of organic cation transporter.
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Release of acetylcholine from murine embryonic stem cells: effect of nicotinic and muscarinic receptors and blockade of organic cation transporter.

机译：从鼠胚胎干细胞释放乙酰胆碱：烟碱和毒蕈碱受体的作用以及对有机阳离子转运蛋白的阻断。

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摘要

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, indicating additional release mechanisms activated upon inhibition of organic cation transporters.Murine embryonic stem cells contain and release significant amounts of acetylcholine. The high fractional release rate and the compensation for blocked organic cation transporters indicate that non-neuronal acetylcholine may play a functional role in the homeostasis of murine embryonic stem cells.
机译：非神经胆碱能系统在自然界广泛表达。进行本实验以表征鼠胚胎干细胞（CGR8细胞系）中的非神经胆碱能系统。将CGR8细胞在装有格拉斯哥缓冲基本必需培养基（Gibco，德国）的糊化烧瓶中培养。通过HPLC结合生物反应器和电化学检测来测量乙酰胆碱.CGR8细胞含有1.08±0.12 pmol乙酰胆碱/ 10（6）个细胞（n = 7），减少为0.50±0.06 pmol / 10（6）个细胞（n = 6;在30μM溴乙酰胆碱存在下（4h）阻断胆碱乙酰基转移酶（p <0.05）。当胆碱酯酶活性被10μM毒扁豆碱阻止时，乙酰胆碱随时间的释放被证明是温育介质，温育期后65×10（6）细胞释放出97±13、180±15和216±14 pmol。分别为2、4和6小时。累积释放对应于2％/ min的分数释放速率。尼古丁或毒蕈碱受体的阻滞没有明显调节乙酰胆碱的释放，乙酰胆碱的释放被300μM奎宁（有机阳离子转运蛋白的抑制剂）大大降低了。这种抑制作用在整个孵育过程中显示出相当程度的褪色，表明在抑制有机阳离子转运蛋白后激活了其他释放机制。小鼠胚胎干细胞含有并释放大量的乙酰胆碱。较高的分数释放速率和对封闭的有机阳离子转运蛋白的补偿表明，非神经元乙酰胆碱可能在鼠胚胎干细胞的体内稳态中发挥功能性作用。

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