title: Pax6-5a promotes neuronal differentiation of murine embryonic stem cells.
abstract: Pax6 genes are highly conserved and important for eye development. Vertebrates predominantly produce two alternatively spliced Pax6 isoforms, Pax6 and Pax6-5a. Pax6-5a differs from Pax6 by the presence of 14 additional amino acids encoded by exon 5a. These additional amino acids occur in the Pax6 paired domain and thus influence its DNA-binding properties. However, little is known about Pax6-5a's physiological functions. Here we establish murine embryonic stem (ES) cell lines in which expression of either the human Pax6 or Pax6-5a isoform is negatively controlled by tetracycline. We report that, in contrast to Pax6 expression, Pax6-5a expression strongly induces ES cells to differentiate into neurons. Moreover, using DNA microarray analysis, we have identified the transcription factor basic helix loop-helix domain containing, class b2 (bHLHb2) in Pax6-5a-expressing ES cells. Our Pax6 isoform-expressing ES cell lines may serve as useful models for identifying Pax6-regulated genes that are important for neurogenesis and/or eye development.
authors.affiliation: Department of Developmental and Regenerative Biology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Nao
authors.initials: N
authors.lastname: Shimizu
authors.forename: Hajime
authors.initials: H
authors.lastname: Watanabe
authors.forename: Junko
authors.initials: J
authors.lastname: Kubota
authors.forename: Jinzhan
authors.initials: J
authors.lastname: Wu
authors.forename: Ryota
authors.initials: R
authors.lastname: Saito
authors.forename: Tadashi
authors.initials: T
authors.lastname: Yokoi
authors.forename: Takumi
authors.initials: T
authors.lastname: Era
authors.forename: Takeshi
authors.initials: T
authors.lastname: Iwatsubo
authors.forename: Takashi
authors.initials: T
authors.lastname: Watanabe
authors.forename: Sachiko
authors.initials: S
authors.lastname: Nishina
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Azuma
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Katada
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nishina
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005136:Eye Proteins / Q000096:biosynthesis / Q000502:physiology*; D018398:Homeodomain Proteins / Q000096:biosynthesis / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D051379:Mice; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000096:biosynthesis / Q000502:physiology*; D010957:Plasmids; D020033:Protein Isoforms / Q000096:biosynthesis / Q000502:physiology; D012097:Repressor Proteins / Q000096:biosynthesis / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013752:Tetracycline
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D020033:Protein Isoforms; D012097:Repressor Proteins; D013752:Tetracycline