Neural tube closure is a morphogenetic process that transforms the neural plate into a neural tube. Throughout this process, contraction of the actin-myosin network is required for apical constriction of the neural plate and maintenance of cell-cell junctions. Fhod3, a member of formin family proteins that mediate nucleation and polymerization of the actin filament, is expressed in the neural tube and heart. In our previous study, we showed that Fhod3 null mouse embryo exhibits cardiac defects and exencephaly, a type of neural tube closure defect wherein the brain is located outside of the skull. However, the mechanism of Fhod3 to regulate neural tube closure is still lacking. Here, we show that Fhod3 is expressed at the lateral wall of the neural tube in the hindbrain. Closure of the neural tube is normally completed by the E9.5, but Fhod3 null embryo shows a persistently open neural tube from the hindbrain/cervical boundary towards the rostral portion. On the apical side of the closing neural plate, loss of Fhod3 disrupts actin-myosin network as well as the cell-cell junction. Taken together, Fhod3 regulates neural tube closure by mediating contraction of actin-myosin network at the cell-cell junction.