title: Reprogramming amacrine and photoreceptor progenitors into retinal ganglion cells by replacing Neurod1 with Atoh7.
abstract: The specification of the seven retinal cell types from a common pool of retina progenitor cells (RPCs) involves complex interactions between the intrinsic program and the environment. The proneural basic helix-loop-helix (bHLH) transcriptional regulators are key components for the intrinsic programming of RPCs and are essential for the formation of the diverse retinal cell types. However, the extent to which an RPC can re-adjust its inherent program and the mechanisms through which the expression of a particular bHLH factor influences RPC fate is unclear. Previously, we have shown that Neurod1 inserted into the Atoh7 locus activates the retinal ganglion cell (RGC) program in Atoh7-expressing RPCs but not in Neurod1-expressing RPCs, suggesting that Atoh7-expressing RPCs are not able to adopt the cell fate determined by Neurod1, but rather are pre-programmed to produce RGCs. Here, we show that Neurod1-expressing RPCs, which are destined to produce amacrine and photoreceptor cells, can be re-programmed into RGCs when Atoh7 is inserted into the Neurod1 locus. These results suggest that Atoh7 acts dominantly to convert a RPC subpopulation not destined for an RGC fate to adopt that fate. Thus, Atoh7-expressing and Neurod1-expressing RPCs are intrinsically different in their behavior. Additionally, ChIP-Seq analysis identified an Atoh7-dependent enhancer within the intronic region of Nrxn3. The enhancer recognized and used Atoh7 in the developing retina to regulate expression of Nrxn3, but could be forced to use Neurod1 when placed in a different regulatory context. The results indicate that Atoh7 and Neurod1 activate distinct sets of genes in vivo, despite their common DNA-binding element.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. cmao@mdanderson.org
authors.forename: Chai-An
authors.initials: CA
authors.lastname: Mao
authors.forename: Jang-Hyeon
authors.initials: JH
authors.lastname: Cho
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Zhiguang
authors.initials: Z
authors.lastname: Gao
authors.forename: Ping
authors.initials: P
authors.lastname: Pan
authors.forename: Wen-Wei
authors.initials: WW
authors.lastname: Tsai
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Frishman
authors.forename: William H
authors.initials: WH
authors.lastname: Klein
mesh_terms: D025042:Amacrine Cells / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D047369:Chromatin Immunoprecipitation; D004596:Electroretinography; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D056426:Genetic Loci; D007150:Immunohistochemistry; D007438:Introns; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D012160:Retina / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; C117638:Atoh7 protein, mouse; C501008:Neurod1 protein, mouse