Cell Adhesion-Dependent Biphasic Axon Outgrowth Elucidated by Femtosecond Laser Impulse
Yamada, S.; Baba, K.; Inagaki, N.; Yoichiroh, H.
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Axon outgrowth is promoted by the mechanical coupling between F-actin and adhesive substrates via clutch and adhesion molecules in an axonal growth cone. In this study, we utilized a femtosecond laser-induced impulse to break the coupling between the growth cone and the substrate, enabling us to evaluate the strength of the binding between the growth cone and a laminin on the substrate, and also determine the contribution of adhesion strength to neurite outgrowth and traction force for the outgrowth. We found that the adhesion strength of axonal L1 cell adhesion molecule (L1CAM)-laminin binding increased with the laminin density on the substrate. In addition, fluorescent speckle microscopy revealed that the retrograde flow of F-actin in the growth cone was dependent on the laminin density such that the flow speed reduced with increasing L1CAM-laminin binding. However, neurite outgrowth and the traction force did not increase monotonically with increased L1CAM-laminin binding but rather exhibited biphasic behavior, in which the outgrowth was suppressed by excessive L1CAM-laminin binding. Our quantitative evaluations suggest that the biphasic outgrowth is regulated by the balance between traction force and adhesion strength. These results imply that adhesion modulation is key to the regulation of neurite outgrowth.
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