Sensitive and Selective Measurement of Hydroxyl Radicals at Subcellular Level with Tungsten Nanoelectrodes
Shushu Ding, Meina Li, Haoyang Gong, Qing Zhu, Guoyue Shi, Anwei Zhu
Abstract
Hydroxyl radical (•OH) is an essential reactive oxygen species involved in critical cell functions. However, the mechanisms controlling its subcellular localization and intracellular level during health and disease remain poorly understood. This is due to the challenge of detecting •OH that are highly reactive and consequently short-lived (in vivo half-life of ∼10–9 s). Herein, we present tungsten nanoelectrodes functionalized with stable 1-hexanethiol (HAT) for selective and sensitive detection of •OH at the subcellular level via the destruction of the self-assembled monolayer of HAT on the nanoelectrode tip. Taking advantage of the ultrasmall nanotip and the super mechanical toughness, the tungsten nanoelectrode could easily penetrate a single living cell without inducing any observable damage. Controlled by a high precision micromanipulator, the •OH level in RAW 264.7 murine macrophages under amyloid β (Aβ) induced oxidative stress were first investigated by the nanoelectrodes at the subcellular level. Moreover, the results revealed the cordycepin-mediated cytoprotection of macrophages through modulation of PI3K/Akt pathway activity and introduction of heme oxygenase-1 (HO-1). We believe that the developed nanoelectrochemical method has shown great capacities for the study of potential drugs for therapeutic intervention of Alzheimer’s disease.