The present study makes an investigation into expression of genes related to cardiac development in chicken, quail and chicken-quail hybrids during the early stage of embryogenesis. Real-time PCR was used to detect mRNA expressions of Nkx2-5, GATA4 and TBX5 in the heart of chicken, quail and chicken-quail hybrids embryos during the 3rd to 7th days of incubation. Results showed that NKX2-5 mRNA displayed a similar expression trend in chicken, quail and chicken-quail hybrids. The initial and highest expression of Nkx2-5 was focused on the 3rd day of incubation, then it declined till 5th day of incubation, thereafter, it fluctuated. Expression of Nkx2-5 gene in quail was significantly higher than in chicken and chicken-quail hybrids, and no significant difference was observed between the two latter species. GATA4 mRNA showed a similar expression trend between chicken and quail, which displayed a steady increase from 3rd to 6th d, then, the expression level decreased. However, GATA4 mRNA expression in chicken-quail hybrids was significantly higher than that in chicken and quail from 3rd to 5th d (p<0.01), but significantly lower than that in chicken and quail during the later stage of the experiment (p<0.05), due to the dramatic drop from 5th d onwards (p<0.01). TBX5 mRNA expression in chicken and quail showed the same trend as GATA4 expressed in the two species. Furthermore, TBX5 expression in chicken-quail hybrids was significantly higher than that in chicken and quail during the whole course of experiment, although relatively lower TBX5 expression was detected in the early stage. In conclusion, Nkx2-5, GATA4 and TBX5 genes showed dynamic changes during the process of cardiac development in chicken, quail and their hybrids embryos. In addition, the expression trend in chicken was similar to that in quail, and there was no significant difference for gene expression level, except NKX2-5. However, expression of these genes in chicken-quail hybrids was significantly different from their parents, the difference mechanism needs to be further explored.