title: Molecular organization of the desmosome as revealed by direct stochastic optical reconstruction microscopy.
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.
authors: Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors: Sara N
authors: SN
authors: Stahley
authors: Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors: Emily I
authors: EI
authors: Bartle
authors: Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA.
authors: Claire E
authors: CE
authors: Atkinson
authors: 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.
authors: Andrew P
authors: AP
authors: Kowalczyk
authors: Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA mattheyses@emory.edu.
authors: Alexa L
authors: 0000-0002-5119-7750
authors: AL
authors: Mattheyses
mesh_terms: D002448:Cell Adhesion; D003896:Desmosomes / Q000378:metabolism*; D006801:Humans; D008297:Male; D008853:Microscopy / Q000379:methods*; D055620:Optical Phenomena; D051191:Plakophilins / Q000378:metabolism; D012867:Skin / Q000378:metabolism; D013269:Stochastic Processes
chemical_list: D051191:Plakophilins
keywords: Cadherin; Cell adhesion; Fluorescence; Keratinocyte; Microscopy; Super-resolution; dSTORM