Gold nanoparticle-assisted all optical localized stimulation and monitoring of Ca2+ signaling in neurons

Abstract

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 Ca2+ 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 Ca2+, measured optically with GCaMP6s. We show that action potentials, measured electrophysiologically, can be induced with this approach. We demonstrate activation of local Ca2+ transients and Ca2+ 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.

Uncontrolled Keywords

Neurons; Membranes; Cells; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Gold; Complement System Proteins; Action Potentials; Light; Lasers; Microscopy, Confocal; Biophysical methods; Ca2+ imaging; Fluorescence imaging; Nanoparticles; Nanoscale biophysics