title: Medaka insulin-like growth factor-2 supports self-renewal of the embryonic stem cell line and blastomeres in vitro.
abstract: Insulin-like growth factors (IGFs) regulate diverse processes including energy metabolism, cell proliferation and embryonic development. They activate the IGF signaling pathway via binding to cell surface receptors. Here we report an essential role of IGF2 in maintaining the pluripotency of embryonic stem (ES) cell from medaka (Oryzias latipes). The medaka igf2 gene was cloned for prokaryotically expression of IGF2 ligand and green fluorescent protein-tagged IGF2 namely IGF2:GFP. With flow cytometry analysis, we demonstrated that the IGF2:GFP can bind to the cultured ES cells from medaka and zebrafish respectively. We also verified that IGF2 is able to activate the phosphorylation of Erk1/2 and Akt, and sustain the viability and pluripotency of medaka ES cells in culture. Furthermore, we characterized the binding of IGF2:GFP to freshly isolated blastomeres by fluorescence microscopy and electron microscopy. Most importantly, we revealed the important role of IGF2 in supporting the derivation of blastomeres in short-term culture. Therefore, Medaka IGF2 is essential for the self-renewal of cultured ES cells and blastomeres from fish embryos. This finding underscores a conserved role of the IGF signaling pathway in stem cells from fish to mammals.
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore, 117543, Singapore. dbshyh@nus.edu.sg.
authors.forename: Yunhan
authors.initials: Y
authors.lastname: Hong
mesh_terms: D000818:Animals; D001757:Blastomeres / Q000166:cytology* / Q000378:metabolism; D000066673:Cell Self Renewal; D002470:Cell Survival; D002478:Cells, Cultured; D003001:Cloning, Molecular; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D007335:Insulin-Like Growth Factor II / Q000235:genetics* / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D008856:Microscopy, Fluorescence; D009990:Oryzias / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D015027:Zebrafish / Q000196:embryology / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D007335:Insulin-Like Growth Factor II