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J Cell Sci. 2016 Aug 1;129(15):2897-904. doi: 10.1242/jcs.185785. Epub 2016 Jun 17.

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

Author information

1
Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
2
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA.
3
Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA Department of Dermatology, Emory University School of Medicine, Atlanta, GA 30322, USA.
4
Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA mattheyses@emory.edu.

Abstract

Desmosomes are macromolecular 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 C-terminal 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.

KEYWORDS:

Cadherin; Cell adhesion; Fluorescence; Keratinocyte; Microscopy; Super-resolution; dSTORM

PMID:
27505428
PMCID:
PMC5004873
DOI:
10.1242/jcs.185785
[Indexed for MEDLINE]
Free PMC Article

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