title: Specification of embryonic stem cell-derived tissues into eye fields by Wnt signaling using rostral diencephalic tissue-inducing culture.
abstract: The eyes are subdivided from the rostral diencephalon in early development. How the neuroectoderm regulates this subdivision, however, is largely unknown. Taking advantage of embryonic stem cell (ESC) culture using a Rax reporter line to monitor rostral diencephalon formation, we found that ESC-derived tissues at day 7 grown in Glasgow Minimum Expression Media (GMEM) containing knockout serum replacement (KSR) exhibited higher levels of expression of axin2, a Wnt target gene, than those grown in chemically defined medium (CDM). Surprisingly, Wnt agonist facilitated eye field-like tissue specification in CDM. In contrast, the addition of Wnt antagonist diminished eye field tissue formation in GMEM+KSR. Furthermore, the morphological formation of the eye tissue anlage, including the optic vesicle, was accompanied by Wnt signaling activation. Additionally, using CDM culture, we developed an efficient method for generating Rax+/Chx10+ retinal progenitors, which could become fully stratified retina. Here we provide a new avenue for exploring the mechanisms of eye field specification in vitro.
authors.affiliation: Laboratory for in vitro Histogenesis, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
authors.forename: Eriko
authors.initials: E
authors.lastname: Sakakura
authors.affiliation: Laboratory for in vitro Histogenesis, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
authors.forename: Mototsugu
authors.initials: M
authors.lastname: Eiraku
authors.affiliation: Laboratory for in vitro Histogenesis, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan; Center for Vascular and Developmental Biology, Feinberg Cardiovascular Research Institute, Northwestern University Feinberg School of Medicine, 303 East Superior Street, Chicago 60611, IL, USA. Electronic address: nozomutakata@cdb.riken.jp.
authors.forename: Nozomu
authors.initials: N
authors.lastname: Takata
mesh_terms: D000818:Animals; D060466:Axin Protein / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D004027:Diencephalon / Q000254:growth & development; D047108:Embryonic Development / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005123:Eye / Q000196:embryology / Q000254:growth & development* / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D020646:Intralaminar Thalamic Nuclei / Q000254:growth & development; D051379:Mice; D012160:Retina / Q000254:growth & development / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D060449:Wnt Signaling Pathway / Q000187:drug effects
chemical_list: D060466:Axin Protein; C111729:Axin2 protein, mouse; D005136:Eye Proteins; D018398:Homeodomain Proteins; C105441:Rax protein, mouse; D014157:Transcription Factors; C088975:Vsx2 protein, mouse
keywords: Embryonic stem cells; Eye field; Optic vesicle; Rax; Rostral diencephalon; Wnt signaling