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Objective Radial migration is critical for the lamination of cerebral cortex during early brain development.Whereas the migration of neurons requires a precise regulation of the organization and dynamics of cytoskeleton including microtubule and actin network, the underlying signaling mechanisms are not well understood.In the present study, we investigate the functional roles of a kinase Mst3 in radial migration during cortical development.Methods The radial migration in mouse neocortex was investigated by in utero electroporation.Growth cone collapse assay in cultured hippocampal neurons was performed using confocal microscopy.Results (1) We identified that Mst3, a serine/threonine kinase highly expressed in the developing mouse brain, regulates the neuronal differentiation.(2) Silencing of Mst3 by in utero electroporation resulted in a significant delay on the neuronal migration in the developing mouse neocortex.(2) Kinase activity of Mst3 was essential for its functions in neuronal morphogenesis and migration.(3) Mst3 was co-localized with F-actin at the growth cones of cultured hippocampal neurons, and regulated actin dynamics.Conclusion Kinase activity of Mst3 is important for neuronal morphogenesis and radial migration through modulation of actin dynamics.