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. 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-term, rod, and C-term) 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 if plaque organization changed in different adhesive states. Plaque-to-plaque distance for DP rod and DP C-term 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 of desmoplakin C-term but not desmoplakin rod or plakoglobin between basal and suprabasal cells. Our data reveal the desmosome molecular organization in cultured keratinocytes and skin as defined by dSTORM.