Browsing by Author "Grooman, Brian"

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    THE ROLE OF THE ACTIN CROSSLINKER PALLADIN: FROM RECONSTITUTED NETWORKS TO LIVE CELLS
    (2013) Grooman, Brian; Upadhyaya, Arpita; Physics; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
    Biophysics is a rapidly growing area of research. New discoveries continue to show the importance of mechanical phenomenon in biological processes, even at the cellular and sub-cellular levels. The complexity of living cells, coupled with their small size makes their study particularly difficult. Palladin is an actin-crosslinker that has not yet been studied as much as other actin-crosslinkers. It localizes with alpha-actinin in stress fibers in many adult cell types. Palladin's exact purpose is still unknown. Through in-vitro studies of reconstituted actin networks we gain insight into the mechanical importance of this novel protein, and show that when partnered with α-actinin, palladin efficiently enhances the network stiffness. Pancreatic Stellate Cells are responsible for maintaining organ integrity, and their malignant counterparts are responsible for one of the most deadly forms of cancer. Interestingly, palladin is shown to be up-regulated in tumors derived from these cells. By studying the stiffness of the cells with and without palladin (via genetic manipulation) we investigate the mechanical importance of palladin in vivo. GFP labelled palladin can serve as a useful marker because it naturally localizes into a regular pattern along stress fibers. Combined with image processing, this makes tracking local strain rates within the cell possible. Pancreatic stellate cells will respond to an applied force by actively contracting their stress fibers. The dynamics of these responses are quantified by tracking the spots of palladin. Through various pharmacological manipulations we study possible signaling pathways that lead from an applied force to stress fiber contraction. Overall, this work explores the mechanical importance of palladin and also investigates the mechanical properties of tumor-associated pancreatic stellate cells, neither of which have been previously studied. Our work shows that palladin controls network stiffness in-vitro, but not in-vivo, suggesting a yet undiscovered purpose. We have also shown that pancreatic stellate cells are in the same range of stiffness as other fibroblasts, and can actively respond to external forces. All of these findings contribute to an increased understanding of the complex systems which govern the mechanical properties of living material.