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Methods Mol Biol. 2013;991:237-47. doi: 10.1007/978-1-62703-336-7_22.

Tracing the endocytic pathways and trafficking kinetics of cell signaling receptors using single QD nanoparticles.

Author information

1
Department of Biomedical Engineering, Oregon Health and Science University, Portland, OR, USA.

Abstract

Cellular signaling is the fundamental process through which cells communicate with each other and respond to their environment. Regulation of this cellular signaling is crucial for healthy cellular function. Malfunctions in signaling are the cause for many diseases and disorders and therefore are under heavy investigation. The molecular mechanisms that underlie cellular signaling rely upon complex and dynamic processes of receptor intracellular trafficking. The specific endosomal pathways and kinetics through which receptors are intracellularly transported regulate the strength and duration of cellular signaling. In even more subtle and complex aspects, the cell orchestrates the individual motions of many receptors, through multiple different pathways, simultaneously. Despite the fundamental role of endosomal trafficking in signal regulation, it has been technically challenging to study since intracellular trafficking is complex and dynamic, with millions of individual receptors simultaneously undergoing trafficking in different endocytic stages. Here, we describe the use of single nanoparticle quantum dot (QD) probes to quantitatively investigate the endocytic trafficking pathways that receptors undergo following ligand activation. This new capability to directly visualize and quantitate cellular signaling at the level of individual receptors inside the cell has broad and important value for understanding fundamental cell signaling processes and the action and effect of therapeutics upon signaling.

PMID:
23546674
DOI:
10.1007/978-1-62703-336-7_22
[Indexed for MEDLINE]

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