Affiliated with Université Laval & CERVO Research Centre

Spatial intensity distribution analysis (SpIDA): a new tool for receptor tyrosine kinase activation and transactivation quantification.

TitleSpatial intensity distribution analysis (SpIDA): a new tool for receptor tyrosine kinase activation and transactivation quantification.
Publication TypeJournal Article
Year of Publication2013
AuthorsBarbeau A, Swift JL, Godin AG, De Koninck Y, Wiseman PW, Beaulieu J-M
JournalMethods Cell Biol
Volume117
Pagination1-19
Date Published2013
ISSN0091-679X
KeywordsApomorphine, Cell Line, Fluorescent Antibody Technique, Gene Expression Regulation, Green Fluorescent Proteins, Humans, Ligands, Microscopy, Confocal, Microscopy, Fluorescence, Molecular Imaging, Neurons, Protein Multimerization, Quinazolines, Receptor, Epidermal Growth Factor, Receptor, trkB, Receptors, Dopamine, Recombinant Fusion Proteins, Software, Transcriptional Activation, Tyrphostins
Abstract

<p>This chapter presents a general approach for the application of spatial intensity distribution analysis (SpIDA) to pharmacodynamic quantification of receptor tyrosine kinase homodimerization in response to direct ligand activation or transactivation by G protein-coupled receptors. A custom graphical user interface developed for MATLAB is used to extract quantal brightness and receptor density information from intensity histograms calculated from single fluorescence microscopy images. This approach allows measurement of monomer/oligomer protein mixtures within subcellular compartments using conventional confocal laser scanning microscopy. Application of quantitative pharmacological analysis to data obtained using SpIDA provides a universal method for comparing studies between cell lines and receptor systems. In addition, because of its compatibility with conventional immunostaining approaches, SpIDA is suitable not only for use in recombinant systems but also for the characterization of mechanisms involving endogenous proteins. Therefore, SpIDA enables these biological processes to be monitored directly in their native cellular environment. </p>

DOI10.1016/B978-0-12-408143-7.00001-3
Alternate JournalMethods Cell Biol.
PubMed ID24143969
Grant List / / Canadian Institutes of Health Research / Canada