Electrophysiological recording of re-aggregating brain cell cultures on multi-electrode arrays to detect acute neurotoxic effects.
Electrophysiological recording of re-aggregating brain cell cultures on multi-electrode arrays to detect acute neurotoxic effects.
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DOI：
10.1016/j.neuro.2007.06.004
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发表时间：
2007-11
期刊：
影响因子：
3.4
通讯作者：
E. van Vliet;L. Stoppini;M. Balestrino;C. Eskes;C. Griesinger;T. Sobański;M. Whelan;T. Hartung;S. Coecke
中科院分区：
文献类型：
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作者：
E. van Vliet;L. Stoppini;M. Balestrino;C. Eskes;C. Griesinger;T. Sobański;M. Whelan;T. Hartung;S. Coecke
神经毒性旨在了解外源性物质如何干扰神经系统的功能并揭示其作用机制。神经元活动是神经系统的主要功能输出，偏离其静息水平可能表明存在毒性。因此，复杂细胞培养系统中电生理活动的监测对于神经毒性评估似乎特别有前景。为了检测化学品的急性神经毒性作用，我们开发了一种测试系统，该系统基于使用多电极阵列重新聚集脑细胞培养物时神经网络的电生理记录。我们表征了培养物的电生理学特性，并使用已知的神经毒物评估了它们在预测神经毒性作用方面的有用性。聚集体表现出诱发场电位和涉及谷氨酸能和 GABA 能突触传递的自发神经活动。配对脉冲抑制表明通过功能性抑制网络存在短期突触可塑性。用 0.1-100 µM 氯化三甲基锡 (TMT)、氯化甲基汞 (MeHgCl)、对硫磷或对氧磷以及 0.1 - 100 mM 乙醇处理培养物长达 100 分钟。 TMT (10 µM)、MeHgCl (1 µM) 和乙醇 (100 mM) 均降低了诱发场电位的幅度。乙醇的作用是可逆的。相比之下，对氧磷 (10 µM) 增加了诱发场电位的幅度，而对硫磷则没有表现出明显的影响。 TMT 和乙醇对自发活动频率的影响与诱发场电位获得的影响一致。所有效应均发生在无法检测到细胞毒性损伤的水平。总的来说，我们的系统表现出与已建立的切片培养制剂相似的电生理特性。它检测到亚细胞毒性水平的已知神经毒物，因此似乎适合评估特别干扰神经系统功能的毒性损伤，例如神经元活动、突触传递和短期可塑性。如果纳入测试策略，它可能是神经毒性作用机制评估的一个有价值的工具。
Neurotoxicity aims to understand how xenobiotics interfere with the function of the nervous system and to unravel their mechanisms of action. Neuronal activity is the primary functional output of the nervous system and deviations from its resting level may indicate toxicity. Consequently, the monitoring of electrophysiological activity in complex cell culture systems appears particularly promising for neurotoxicity assessment. To detect acute neurotoxic effects of chemicals we developed a test system based on the electrophysiological recordings from neural networks in re-aggregating brain cell cultures using multi-electrode arrays. We characterised the electrophysiological properties of the cultures and, using known neurotoxicants, evaluated their usefulness to predict neurotoxic effects. Aggregates displayed evoked field potentials and spontaneous neural activity involving glutamatergic and GABAergic synaptic transmission. Paired pulse inhibition indicated the presence of short-term synaptic plasticity via functional inhibitory networks. Cultures were treated with 0.1 100 µM of trimethyltin chloride (TMT), methyl mercury chloride (MeHgCl), parathion or paraoxon, and with 0.1 - 100 mM of ethanol for up to 100 minutes. TMT (10 µM), MeHgCl (1 µM) and ethanol (100 mM) all decreased the amplitude of evoked field potential. The effect of ethanol was reversible. In contrast paraoxon (10 µM) increased the amplitudes of evoked field potentials while parathion showed no significant effects. The effects of TMT and ethanol on the frequency of spontaneous activity were consistent with those obtained for evoked field potentials. All effects occurred at levels at which cytotoxic injuries were not detectable. Taken together our system expressed electrophysiological properties similar to those of established slice culture preparations. It detected known neurotoxicants at subcytotoxic levels and therefore appears suitable for the assessment of toxic insults specifically interfering with nervous system function, e.g. neuronal activity, synaptic transmission and short-term plasticity. If incorporated into testing strategies, it might represent a valuable tool for the mechanistic assessment of neurotoxic effects.