Angiogenesis is the process of new vasculature formation from existing vessels, using cues from the extracellular matrix (ECM) to stimulate endothelial cells (ECs) to extend branches, polarize, and migrate. However, the contribution of microtubules (MTs) to the processes to promote directed cell migration are not well known. MTs play an essential role in determining the branching patterns and directionality of the ECs to their target locations. The localized MT growth instability of MTs is dependent on mitotic centromere-associated kinesin (MCAK) in cells during interphase. Polarized cells exhibit decreased MCAK activity at the leading edge and thus increased MT growth there compared to the trailing edge. In addition to their structural role in defining EC morphology, MTs also function as intracellular roadways along which molecular motor proteins traffic cargo. MMP14 is a transmembrane zinc metalloprotease that is trafficked on MTs to the cell surface where it modifies and degrades the ECM. It is not known how MMP14 trafficking to the cell surface is regulated by the dynamics of MTs or how MMP14 activity is controlled with local specificity at the cell surface. The ability to study MT growth dynamics using live-cell imaging is an invaluable tool as we study the movement of MMP14 to the cell surface. Coupling this with the ability to monitor MMP14 activity in living cells as they migrate through a collagen-rich matrix is especially useful. To date, there exists no plasmid–based biosensor that can be used as a readout of MMP activity. We have developed an MMP14 biosensor that will allow us to elucidate and bring together the mechanisms of how MTs aid in polarizing cells together with MMP14 and promote directed cell migration. This work elucidated the cellular mechanisms by which localized MT dynamic instability was affected before cell polarization and developed a biosensor to study how MMP14 activity in these polarized affects cell migration.
|School:||University of the Sciences in Philadelphia|
|Department:||Cell and Molecular Biology|
|School Location:||United States -- Pennsylvania|
|Source:||DAI-B 80/08(E), Dissertation Abstracts International|
|Subjects:||Molecular biology, Cellular biology|
|Keywords:||Cell migrate, Lice-cell imaging, Microtubules, Mlak, Mmp14, Racl|
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