The Role of Myosin 2 in Contact Guidance

National Heart, Lung, And Blood Institute

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Abstract

To investigate the contribution of NMs 2 to cell contact guidance we established a microprinting technique. We tested the effect of the NM 2B isoform knockout by visualizing cell spreading and dynamics of the cytoarchitecture on fibronectin coated micro-patterns on soft acrylamide gels and hard polydimethylsiloxane surfaces. This allows us to elucidate proteins that guide spreading and migration of fibroblast cells. We found that absence of NM 2B altered contact guidance of fibroblasts manifesting in a substantial loss of cellular projections and uniaxial directional polarization as well as increased contractility and elasticity of cells. Using cytoskeleton targeting compounds and a series of genetic transformations we verified a possible role for the NM 2B isoform in contact guidance and proposed a model in which the roles of NM 2A and 2B isoforms diverge in control over structure of the actin cytoskeleton and its crosstalk with microtubules. To assess reproducibility and verify the role of NMs 2 in contact guidance, an independent series of cell lines will be tested. This second series will accommodate treatment with cytoskeleton targeting compounds and a set of genetic transformation to affect gene expression in a manner similar to the one used previously for fibroblast cells. We are creating and testing a model of cell cytoskeleton architecture dynamics that incorporates actin-tubulin-myosin dependent regulation of contact guidance. This model creates a platform for the study of the roles of NMs 2 in tissue development.