Multipolar-to-bipolar transition (MBT) of pyramidal cell is a vital prerequisite for both radial glia-guided migration and axon-dendrite polarity in the developing cerebral cortex. However, the molecular and/or cellular mechanisms spatiotemporally orchestrating MBT remain unknown. Here we report that N-cadherin-mediated radial glial cell-cortical neuron interaction directs axon formation from the opposite side of contacting area in vitro. In vivo, N-cadherin is accumulated at the binding site between radial glial fiber and leading-like process of unpolarized pyramidal cell and inhibition of these N-cadherin-mediated adhesions in pyramidal cell disrupts its MBT. Furthermore, MBT zone localizes between the upper and lower intermediate zone (IZ) and it notably coincides with the appearance of predominant expression of N-cadherin in the upper IZ. Finally, we show that N-cadherin-mediated radial glial cell-cortical neuron interaction directs axon formation through activating RhoA in the contacting neurite and localizing activated Rac in the opposite neurite. Pyramidal cells expressing RhoA-DN show a similar phenotype to inhibition of N-cadherin in pyramidal cell and fail to transit from multipolar stage to bipolar stage. Together, these data suggest a novel model in which N-cadherin-mediated radial glial cell-pyramidal cell interaction directs MBT.