Lavoie-Cardinal Flavie, Salesse Charleen, Bergeron Éric, Meunier Michel, De Koninck Paul
Institut Universitaire en Santé Mentale de Québec, 2601 de la Canardière, Québec, QC, G1J 2G3, Canada.
Laser Processing and Plasmonics Laboratory, Engineering Physics Department, École Polytechnique de Montréal, Montréal, QC, H3C 3A7, Canada.
Sci Rep. 2016 Feb 9;6:20619. doi: 10.1038/srep20619.
Light-assisted manipulation of cells to control membrane activity or intracellular signaling has become a major avenue in life sciences. However, the ability to perform subcellular light stimulation to investigate localized signaling has been limited. Here, we introduce an all optical method for the stimulation and the monitoring of localized Ca(2+) signaling in neurons that takes advantage of plasmonic excitation of gold nanoparticles (AuNPs). We show with confocal microscopy that 800 nm laser pulse application onto a neuron decorated with a few AuNPs triggers a transient increase in free Ca(2+), measured optically with GCaMP6s. We show that action potentials, measured electrophysiologically, can be induced with this approach. We demonstrate activation of local Ca(2+) transients and Ca(2+) signaling via CaMKII in dendritic domains, by illuminating a single or few functionalized AuNPs specifically targeting genetically-modified neurons. This NP-Assisted Localized Optical Stimulation (NALOS) provides a new complement to light-dependent methods for controlling neuronal activity and cell signaling.
利用光辅助操纵细胞来控制膜活性或细胞内信号传导已成为生命科学的一条主要途径。然而，进行亚细胞光刺激以研究局部信号传导的能力一直受到限制。在此，我们介绍一种全光学方法，用于刺激和监测神经元中的局部Ca(2+)信号传导，该方法利用了金纳米颗粒（AuNPs）的等离子体激发。我们通过共聚焦显微镜显示，将800纳米激光脉冲施加到装饰有少量AuNPs的神经元上会触发游离Ca(2+)的瞬时增加，这是用GCaMP6s进行光学测量的。我们表明，通过这种方法可以诱导通过电生理学测量的动作电位。我们通过照亮专门靶向基因修饰神经元的单个或少数功能化AuNPs来证明在树突域中通过CaMKII激活局部Ca(2+)瞬变和Ca(2+)信号传导。这种NP辅助局部光学刺激（NALOS）为控制神经元活动和细胞信号传导的光依赖方法提供了一种新的补充手段。
