Journal article

Fluorescent Labeling and Quantification of Vesicular ATP Release Using Live Cell Imaging

Kirstan A Vessey, Tracy Ho, Andrew I Jobling, Anna Y Wang, Erica L Fletcher, P Pelegrin (ed.)

Methods in molecular biology (Clifton, N.J.) | HUMANA PRESS INC | Published : 2020

Abstract

Adenosine triphosphate (ATP) is actively transported into vesicles for purinergic neurotransmission by the vesicular nucleotide transporter, VNUT, encoded by the gene, solute carrier 17, member 9 (SLC17A9). In this chapter, methods are described for fluorescent labeling of VNUT positive cells and quantification of vesicular ATP release using live cell imaging. Directions for preparation of viable dissociated neurons and cellular labeling with an antibody against VNUT and for ATP containing synaptic vesicles with fluorescent ATP markers, quinacrine or MANT-ATP, are detailed. Using confocal microscope live cell imaging, cells positive for VNUT can be observed colocalized with fluorescent ATP v..

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