Luo, D., Hu, W., Chen, S., Xiao, Y., Sun, Y., and Zhu, Z. (2009) Identification of Differentially Expressed Genes Between Cloned and Zygote-Developing Zebrafish (Danio rerio) Embryos at the Dome Stage Using Suppression Subtractive Hybridization. Biology of reproduction. 80(4):674-684.
Comparative analyses of differentially expressed genes between somatic cell nuclear transfer (SCNT) embryos and zygote-developing (ZD) embryos are important for understanding the molecular mechanism underlying the reprogramming processes. Herein, we employed the suppression subtractive hybridization (SSH) approach and from more than 2,900 clones identified 96 differentially expressed genes between the SCNT and ZD embryos at the dome stage in zebrafish. We have reported the first database of differentially expressed genes in zebrafish SCNT embryos. Collectively, our findings demonstrate that zebrafish SCNT embryos undergo significant reprogramming processes during the dome stage. However, the majority of differentially expressed genes are down-regulated in SCNT embryos, indicating failure of reprogramming. Based on EnsEMBL description and Gene Ontology (GO) annotation, the problems of reprogramming at the dome stage may occur during nuclear remodeling, translation initiation, and regulation of the cell cycle. The importance of regulation from recipient oocytes in cloning should not be underestimated in zebrafish.