Precisely controlled proliferation and differentiation is critical for correct patterning in central nervous system development.However, the regulatory mechanism for balancing proliferation and differentiation remains unclear.Dorsal midbrain, one of the fastest growing tissues, is an excellent platform to investigate this question.Stathmins are a well-known microtubule destabilizing proteins, by modulating microtubule polymerization.They are involved in neurogenesis in culture cells, but their roles during brain development remains illusive.By whole-mount in situ hybridization assay, we observed that specific expression of stathmin-like 4 (stmn4) in the developing zebrafish dorsal midbrain, but not other stathmin isoforms.Using different loss of function approaches, including antisense morpholino,dominant-negative gene expression and shRNA, we found that consistent premature neuronal differentiation in dorsal midbrain.Furthermore, the G2 phase was lengthen or even arrested in dorsal midbrain of stmn4 morphants.Lastly, Wnt signaling, a well known regulator of dorsal midbrain development, was required for stmn4 expression.The forced expression of Wnt antagonist, DKK1, reduced the repression of stmn4 expression at dorsal midbrain.It implies that the stmn4 expression may be mediated directly or indirectly by upstream Wnt signaling.Collectively, our results demonstrate stmn4 plays a crucial role to maintain neuronal cells at progenitor cell state in dorsal midbrain under the control of Wnt signaling and a proper inductive signal then releases the stmn4 brake that allows neuronal differentiation and further development.