Objective The aim of this study was to explore the effect of Pitx2 knock-down on the early tooth development related genes in Zebrafish by Pitx2-morpholino oligos microinjection.Methods Pitx2 gene was kncked down by morpholino oligos microinjection.We Observed the development of Zebrafish by microscopes in 72h,and obtain the RNA of the embryos after microinjection.Then determinate the changes of the expression of Bmp4 and Fgf8 which are involved in tooth development by semi-quantitative RT-PCR and grayscale value analysis.We synthesized gene probes for Bmp4、Fgf8 by cloning specific gene fragment of Bmp4 and Fgf8 with RT-PCR directly,and then carried out the in situ hybridization with embryos from different stages during early dental development to observe the effect of Pitx2 knock-down on the Expression of Bmp4 and Fgf8 in Zebrafish early dental development.Results We synthesized gene probes for Bmp4 and Fgf8 by cloning Bmp4 and Fgf8,and got the pictures and data during early dental development in Zebrafish.The embryos appeared to growth retardation or termination,.pericardial edema,spinal and eyes deformity in Morpholino groups.Semi-quantitative RT-PCR and grayscale value analysis indicatesd Pitx2 inhibition increased the expression of Bmp4 and repressed the expression of Fgf8.In situ hybridization showed that Pitx2 knock-down increased the expression of Bmp4,and repressed the expression in branchial arch of Zerafish.Conclusion Down-regulated Pitx2 made Zerafish growing with retardation or yermination,and increased the expression of Bmp4 and repressed the expression of Fgf8.