Cell-to-Cell Connectivity Assays for the Analysis of Cytoskeletal and Other Regulators of Plasmodesmata.

The actin cytoskeleton has close but so far incompletely understood connections to plasmodesmata, the cell junctions of plants. Plasmodesmata are essential for plant development and responses to biotic and abiotic stresses and facilitate the intercellular exchange of metabolites and hormones but also macromolecules such as proteins and RNAs. The molecular size exclusion limited of plasmodesmata is dynamically regulated, including by actin-associated proteins. Therefore, experimental analysis of plasmodesmal regulation can be relevant to plant cytoskeleton research. This chapter presents two simple imaging-based protocols for analyzing macromolecular cell-to-cell connectivity in leaves.