The role of plectin in processes associated with focal adhesions in mouse astrocytesFurlani, Borut (Avtor) Jorgačevski, Jernej (Mentor) plectinastrocytesfocal adhesionsintermediate filamentsreactive astrogliosisPlectin is a cytoskeletal linker protein that is highly enriched in glial cells. This thesis investigates the role of plectin in focal adhesion (FA)-associated processes in cultured mouse astrocytes. Super-resolution microscopy revealed that plectin localizes to FAs, where it exhibits a distinct structural organization and becomes progressively enriched during FA maturation. Live-cell imaging demonstrated that plectin regulates FA turnover during astrocyte spreading and influences the mobility of individual FA proteins. Consistent with the importance of FA dynamics for cell spreading and migration, plectin-deficient astrocytes exhibited reduced single-cell mobility, decreased spreading area and a rounder morphology compared with wild-type astrocytes. FA-cytoskeleton colocalization analyses revealed that loss of plectin reduces the association of FAs with the intermediate filament (IF) protein vimentin. Connectivity analysis showed that plectin deficiency disrupts the architecture of IF and microtubule networks. These alterations were accompanied by decreased cellular stiffness. Given the interaction between plectin and aquaporin 4, the role of plectin in astrocyte volume regulation was examined, revealing altered kinetics of local peripheral swelling. Astrocytes respond to pathological stimuli through reactive astrogliosis, which is characterized by cytoskeletal remodeling and upregulation of IFs. In a model of reactive astrocytes we have confirmed elevated plectin levels along with an increased number and size of FAs, suggesting the involvement of plectin in pathological conditions. Collectively, this work identifies plectin as a regulator of cytoskeletal organization, FA dynamics, cellular mechanics, and local volume regulation in astrocytes.[B. Furlani]20262026-06-23 13:09:00Doktorsko delo/naloga183990sl