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Super-resolution imaging as a method to study GPCR dimers and higher-order oligomers

Jonas, KC; Hanyaloglu, AC (2018) Super-resolution imaging as a method to study GPCR dimers and higher-order oligomers. Neuromethods, 140. pp. 329-343. ISSN 1940-6045 https://doi.org/10.1007/978-1-4939-8576-0_21
SGUL Authors: Jonas, Kim Carol

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Abstract

The study of G protein-coupled receptor (GPCR) dimers and higher-order oligomers has unveiled mechanisms for receptors to diversify signaling and potentially uncover novel therapeutic targets. The functional and clinical significance of these receptor–receptor associations has been facilitated by the development of techniques and protocols, enabling researchers to unpick their function from the molecular interfaces, to demonstrating functional significance in vivo, in both health and disease. Here we describe our methodology to study GPCR oligomerization at the single-molecule level via super-resolution imaging. Specifically, we have employed photoactivated localization microscopy, with photoactivatable dyes (PD-PALM) to visualize the spatial organization of these complexes to <10 nm resolution, and the quantitation of GPCR monomer, dimer, and oligomer in both homomeric and heteromeric forms. We provide guidelines on optimal sample preparation, imaging parameters, and necessary controls for resolving and quantifying single-molecule data. Finally, we discuss advantages and limitations of this imaging technique and its potential future applications to the study of GPCR function.

Item Type: Article
Additional Information: This is a post-peer-review, pre-copyedit version of a chapter published in Receptor-Receptor Interactions in the Central Nervous System. The final authenticated version is available online at: http://dx.doi.org/10.1007/978-1-4939-8576-0_21
SGUL Research Institute / Research Centre: Academic Structure > Institute of Medical & Biomedical Education (IMBE)
Academic Structure > Institute of Medical & Biomedical Education (IMBE) > Centre for Biomedical Education (INMEBE)
Journal or Publication Title: Neuromethods
ISSN: 1940-6045
Dates:
DateEvent
2 August 2018Published
13 August 2018Accepted
Publisher License: Publisher's own licence
Projects:
Project IDFunderFunder ID
BB/1008004/1Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
UNSPECIFIEDSociety for Endocrinologyhttp://dx.doi.org/10.13039/501100000382
URI: https://openaccess.sgul.ac.uk/id/eprint/110048
Publisher's version: https://doi.org/10.1007/978-1-4939-8576-0_21

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