Elucidation Of Arpin Polarisation In Cells Residing On Adhesive Micropatterned Surfaces
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Date
2015-08
Authors
Nageswaran, Mageswary
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Publisher
Universiti Sains Malaysia
Abstract
Cell adhesion and spreading are important components of cell migration. Multiple factors such as organisation of highly dynamic actin filaments and stable lamelipodium formation is crucial for focal adhesion and cell spreading. HeLa cell was used as a cell model. Micropatterns with four shapes (I,Y,crossbow,disk) coated with fibronectin were used to investigate the effects of geometric cues and topography of extracellular matrix protein on cell adhesion and morphology. Cells seeded on uncoated normal glass coverslip act as control. We hypothesised that the geometry of surfaces influences the internal organisation of the cells and cells can change their shape in response to the mechanical forces. The study depicts that micropatterns orchestrate regular cell shape as compared to an irregular shape adopted by the cells grown on uncoated glass cover slip. The assembly of actin filaments and actomyosin stress fibers differs with patterns. To support this data, cell area was obtained to examine the cell spreadiness. As expected, cells on confined geometry (I,Y,crossbow) spread less as compared to the disk-shaped pattern. Cells on disc shaped surface adopted a round shape and randomly polarised. Focal adhesion and membrane protrusion span the periphery. Cells were strongly polarised and adopted the shape of oriental fan on crossbow pattern. On surface with I shape, the cells were adopted pseudo-square shape. Cells on Y shape took up a triangular shape with actomyosin contraction at the three edges whereas actin filament formed at the adhesive edges. The extent of cell spreading is correlated with the total space covered with ECM-proteins. Arpin is an inhibitor of actin nucleation. Thus, the study investigates the polarisation and localisation of Arpin in cells residing on both patterned and non-patterned surfaces. The data obtained suggests that Arpin localises with stress fibers. The results presented illustrates that the presence of Arpin influences actin filament formation thus restrics the protrusion of lamellipodium. RhoA protein plays a key role in actomyosin contractility. Thus, we studied the gene expression of RhoA gene in HeLa cell and found that the cells on patterned surface have a lower expression of RhoA gene compared to expression of RhoA gene expressed by cells grown on uncoated cover slip.
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Keywords
Cell spreading