Syntaxin is a multi-domain protein that is specifically distributed in the presynaptic membrane of neuron. It is one of the key proteins in membrane fusion. However, the roles of Syntaxin in nerve cell differentiation remain unclear. In this study, wild type and different mutats of Syntaxin were overexpressed in the mouse neuroblastoma cells (N2a). Parameters such as differentiation rate of the cell, number of branches and total length of the neurite were analyzed to determine the effects of Syntaxin in neuronal differentiation. Our results suggested the Habc domain of Syntaxin was the main functional structure in differentiation process. The number of branches and total length of the neurite, but not the differential rate, were affected by Syntaxin overexpression.