Characterizing the Effects of Nesprin 2G and Alpha4Beta1 Integrin on the Intracellular Dynamics of Cells in a Migration Model

Cell migration is an important cellular process which has been shown to play a vital role in a variety of phenomena, including embryonic development, tissue healing and the inflammatory response, as well as in pathological conditions such as vascular disease, osteoporosis, chronic inflammatory disease, multiple sclerosis, and most notably in metastatic cancer. Mechanically, migrating cells undergo five major steps: morphological polarization, membrane extension, formation of cell-substratum attachments, contractile force and traction, and release of attachments. This process involves the organization and dynamic activity of both the nucleus and the cytoskeleton in order to ensure proper morphological change. In order to better study this process, a migration model using micropatterns of different s was developed to force cells into different migration-like configurations. This was done to provide a standardized way to quantitatively examine different structural properties involved in cell migration. To that end, this study examined two different proteins, nesprin 2G and α4β1 integrin, and attempted to characterize and measure how they affect the organization of both the cytoskeleton and the nucleus in two different cell lines, Chinese hamster ovary cells and C2C12 mouse myoblasts. Our results indicated that nesprin 2G plays an important role in the organization of both the nucleus and actin stress fibers of C2C12 cells, helping to regulate the orientation of both in migration-like conditions. Likewise, our results indicated that α4β1 integrin plays an important role in the orientation and coherency of actin stress fibers in different morphologies, as well as indirectly affecting the shape and size of the nucleus of CHO cells. Overall, this study showed the potential usefulness of the micropattern migration model and helped to further characterized and demonstrate the function of nesprin 2G and α4β1 integrin.

Barbera, Nicolas

Johns Hopkins University



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