Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy

• Type of Material: Article
• Authors: Sarah N. Stahley, Emory University; Emily I. Bartle, Emory University; Claire E. Atkinson, University of Chicago; Andrew Kowalczyk, Emory University; Alexa Mattheyses, Emory University
• Language: English
• Date: 2016-08-01
• Publisher: Company of Biologists
• Publication Version: Final Published Version
• Copyright Statement: © 2016. Published by The Company of Biologists Ltd
• Final Published Version (URL): http://dx.doi.org/10.1242/jcs.185785
• Title of Journal or Parent Work: Journal of Cell Science
• ISSN: 0021-9533
• Volume: 129
• Issue: 15
• Start Page: 2897
• End Page: 2904
• Grant/Funding Information: This project was supported by the National Institutes of Health [grant numbers R21AR066920 to A.L.M., R01AR048266 to A.P.K.]; and National Science Foundation [grant number IDBR 1353939 to A.L.M.]. SIM was supported by the Integrated Cellular Imaging Core at Emory University.
• Keywords: PEMPHIGUS; MAP; NUCLEAR-PORE COMPLEX; ADHESIVE; Keratinocyte; Cell Biology; Microscopy; FLEXIBILITY; PERSPECTIVE; DESMOPLAKIN; Super-resolution; Science & Technology; Cadherin; Fluorescence; dSTORM; Life Sciences & Biomedicine; ARCHITECTURE; DISEASE; PLAQUE; Cell adhesion
• Research Categories: Biology, Cell; Chemistry, Biochemistry

Abstract

Desmosomes aremacromolecular junctions responsible for providing strong cell-cell adhesion. Because of their size and molecular complexity, the precise ultrastructural organization of desmosomes is challenging to study. Here, we used direct stochastic optical reconstruction microscopy (dSTORM) to resolve individual plaque pairs for inner and outer dense plaque proteins. Analysis methods based on desmosomal mirror symmetry were developed to measure plaque-to-plaque distances and create an integrated map. We quantified the organization of desmoglein 3, plakoglobin and desmoplakin (N-terminal, rod and C-terminal domains) in primary human keratinocytes. Longer desmosome lengths correlated with increasing plaque-to-plaque distance, suggesting that desmoplakin is arranged with its long axis at an angle within the plaque. We next examined whether plaque organization changed in different adhesive states. Plaque-to-plaque distance for the desmoplakin rod and C-terminal domains decreased in PKP-1-mediated hyperadhesive desmosomes, suggesting that protein reorganization correlates with function. Finally, in human epidermis we found a difference in plaque-to-plaque distance for the desmoplakin Cterminal domain, but not the desmoplakin rod domain or plakoglobin, between basal and suprabasal cells. Our data reveal the molecular organization of desmosomes in cultured keratinocytes and skin as defined by dSTORM.