Jump to main content
Jump to site search

Issue 11, 2017
Previous Article Next Article

Quantitative image cytometry for analyzing intracellular trafficking of G protein-coupled receptors on a chemical-trapping single cell array

Author affiliations

Abstract

G protein-coupled receptors (GPCRs) are important targets in medical and pharmaceutical research fields, because they play key roles in a variety of biological processes. Recently, intracellular trafficking of GPCRs involving endosomal internalization and recycling to the plasma membrane has been studied as a regulation mechanism for GPCR activities. However, the absence of a quantitative single-cell analysis method has hampered conditional GPCR trafficking studies and the possibility of gaining significant insights into the mechanism of regulation of GPCR signaling. Here, we report a facile image cytometry method to analyze the trafficking of GPCRs. In this method, GPCR-expressing cells were arrayed with a photo-responsive cell-immobilizing reagent in a single-cell manner, and the tagged GPCR was visualized by pulse-labeling with a fluorescent dye through sortase-mediated peptide-tag ligation. We quantified the intracellular distribution changes of a pH-dependent GPCR, G2A, by time-course observation under mildly acidic and slightly basic pH conditions. The difference in pH-dependent G2A trafficking between individual cells was automatically detected by an image analysis custom software program, and simultaneously, the average distribution ratios were also determined for understanding the properties of G2A. The present method should be applicable for investigating the dynamic intracellular trafficking of a wide variety of GPCRs under various conditions in a high-throughput manner.

Graphical abstract: Quantitative image cytometry for analyzing intracellular trafficking of G protein-coupled receptors on a chemical-trapping single cell array

Back to tab navigation

Supplementary files

Publication details

The article was received on 26 Feb 2017, accepted on 17 Apr 2017 and first published on 18 Apr 2017


Article type: Communication
DOI: 10.1039/C7LC00198C
Citation: Lab Chip, 2017,17, 1933-1938
  •   Request permissions

    Quantitative image cytometry for analyzing intracellular trafficking of G protein-coupled receptors on a chemical-trapping single cell array

    M. Tan, S. Yamaguchi, S. Yamahira, M. Nakamura and T. Nagamune, Lab Chip, 2017, 17, 1933
    DOI: 10.1039/C7LC00198C

Search articles by author

Spotlight

Advertisements