Studying endosomes in cultured neurons by live-cell imaging

Abstract

Endosomes play critical roles on regulating surface receptor levels as well as signaling cascades in all cell types, including neurons. Endocytosis and endosomal trafficking is routinely studied after fixation, but live imaging is increasingly being used to capture the dynamic nature of endosomes and is allowing increasingly sophisticated glimpses into trafficking processes in live neurons. In this chapter, we describe the basics of neuronal primary cultures, methods for expressing fluorescent proteins, and live imaging of cargos and endosomal regulators.

Keywords: Endocytic cargo; Endocytosis; Endosome dynamics; Internalization; Live imaging; Live imaging chamber; Neuronal culture; Primary neurons; Pulse-chase experiments; Rab proteins.