The development of the vertebrate face is regulated by complex interactions among several signaling pathways. Dicldcopf-1 (Dkk-1), an inhibitor of the Wnt/beta-catenin signaling pathway, can affect midfacial morphogenesis. The downstream target genes of the Wnt/beta-catenin signaling pathway in morphogenesis of the developing upper jaw and lip remain unknown. To investigate the functional roles of Wnt/beta-catenin signaling in facial development, we performed a loss-of-function experiment using local implantation of beads soaked with Dkk-1 during lip fusion at the maxillary prominence of chick embryos at stage 22(HH22). Antagonism of Wnt/beta-catenin signaling by Dkk-1 induced deformities of the premaxilla and jugal bone, which are derived from the maxillary mesenchyme. Real-time and semi-quantitative RT-PCR analysis showed the significant reduction of Lhx8, Msx1 and Msx2 expression levels around the beads in the maxillary mesenchyme at 6 and 24 h after bead implantation. Time course experiments in the HH 22 embryos showed the effect of Dkk-1 on Lhx8, Msxl and Msx2 expression was not significant after 48 h of the treatment. At HH 26 when the fusion of facial primordial started, Dkk-1 application did not exhibit any significant reduction of those genes. Our findings suggested that Dkk-1 regulates maxillary morphogenesis in chick embryos through Lhx8, Msxl and Msx2 signals. Wnt/beta-catenin signaling is responsible for intrinsic upper jaw development before the lip fusion. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.