Identification Of Pluripotency Genes In The Fish

Stem cells are pluripotent cells with the ability to self-replicate,which can be differentiated into various cell types.Pluripotency genes are routinely used to characterize mammalian stem cell cultures at the molecular level.Mammalian pluripotency-related factors mainly include transcription factors such as Pou5f1,Oct4,Sox2,Klf4 and Nanog.These genes can not only maintain the pluripotency of stem cells,but also can activata the gene reprogramming in somatic cells by overexpressing.However,such genes have remained unknown in lower vertebrates.Therefore,16 genes that maybe related to pluripotency were collected through literature review.Zebrafish and red crucian carp were used as the research object.The expression characteristics of 16 candidate genes in embryonic and adult tissues were analyzed by RT-PCR.Our result showed that Oct4,Nanog and Gdf3 were highly expressed in early embryos and lowly in the last stage of gastrula.qRT-PCR analysis showed that these genes have maternal expression,high expression in the early stage of blastocyst,and expression levels after differentiation in somite significantly down-regulated.RT-PCR detection also showed that Oct4 and Nanog were specifically expressed in refined gonad,Gdf3 was specifically expressed in refined gonad and brain.RNA probe in situ hybridization localization analysis showed that Oct4,Nanog and Gdf3 were specifically expressed in refined ovarian tissue-derived cells andearly developmental meiocytes.In summary,Oct4,Nanog and Gdf3 can be used as stable fish pluripotency marker gene.This study laid the foundation for the further research on fish stem cells.