Department of Drug Science and Technology, "NIS" inter-departmental centre, University of Torino, Corso Raffaello 30, 10125, Torino, Italy.
Istituto Nazionale Ricerca Metrologica, Strada delle Cacce 91, 10135, Torino, Italy.
Sci Rep. 2018 Feb 2;8(1):2221. doi: 10.1038/s41598-018-20528-5.
Fluorescent nanodiamonds (FND) are carbon-based nanomaterials that can efficiently incorporate optically active photoluminescent centers such as the nitrogen-vacancy complex, thus making them promising candidates as optical biolabels and drug-delivery agents. FNDs exhibit bright fluorescence without photobleaching combined with high uptake rate and low cytotoxicity. Focusing on FNDs interference with neuronal function, here we examined their effect on cultured hippocampal neurons, monitoring the whole network development as well as the electrophysiological properties of single neurons. We observed that FNDs drastically decreased the frequency of inhibitory (from 1.81 Hz to 0.86 Hz) and excitatory (from 1.61 to 0.68 Hz) miniature postsynaptic currents, and consistently reduced action potential (AP) firing frequency (by 36%), as measured by microelectrode arrays. On the contrary, bursts synchronization was preserved, as well as the amplitude of spontaneous inhibitory and excitatory events. Current-clamp recordings revealed that the ratio of neurons responding with AP trains of high-frequency (fast-spiking) versus neurons responding with trains of low-frequency (slow-spiking) was unaltered, suggesting that FNDs exerted a comparable action on neuronal subpopulations. At the single cell level, rapid onset of the somatic AP ("kink") was drastically reduced in FND-treated neurons, suggesting a reduced contribution of axonal and dendritic components while preserving neuronal excitability.
荧光纳米金刚石(FND)是一种基于碳的纳米材料,能够高效地掺入光学活性的光致发光中心,如氮空位复合物,因此它们有望成为光学生物标记物和药物递送剂。FND 具有明亮的荧光,无光漂白,同时具有高摄取率和低细胞毒性。本研究关注 FND 对神经元功能的干扰,研究了其对培养海马神经元的影响,监测整个网络的发育以及单个神经元的电生理特性。结果发现,FND 显著降低了抑制性(从 1.81Hz 降至 0.86Hz)和兴奋性(从 1.61Hz 降至 0.68Hz)微小突触后电流的频率,并通过微电极阵列一致降低动作电位(AP)放电频率(36%)。相反,爆发同步性得以保留,自发性抑制和兴奋性事件的幅度也得以保留。电流钳记录显示,具有高频(快棘波)AP 串的神经元与具有低频(慢棘波)AP 串的神经元的比例没有改变,表明 FND 对神经元亚群具有相似的作用。在单细胞水平上,FND 处理后的神经元体细胞 AP 的起始“扭结”明显减少,提示轴突和树突成分的贡献减少,而神经元兴奋性得以保留。