CoChat
Sign Up
Cited by 158Retinal Development and DisordersDevelopmental Biology and Gene RegulationOcular Disorders and Treatments

A regulatory loop involving PAX6, MITF, and WNT signaling controls retinal pigment epithelium development

Open access
Bharti, Kapil, Gasper, Melanie, Ou, Jingxing, Brucato, Martha, Clore-Gronenborn, Katharina, Pickel, James, Arnheiter, Heinz·2012·PLoS Genetics
DOI 10.1371/journal.pgen.1002757PMC PMC3390378
Open this paper in CoChat — free accountOpen PDF

Abstract

The separation of the optic neuroepithelium into future retina and retinal pigment epithelium (RPE) is a critical event in early eye development in vertebrates. Here we show in mice that the transcription factor PAX6, well-known for its retina-promoting activity, also plays a crucial role in early pigment epithelium development. This role is seen, however, only in a background genetically sensitized by mutations in the pigment cell transcription factor MITF. In fact, a reduction in Pax6 gene dose exacerbates the RPE-to-retina transdifferentiation seen in embryos homozygous for an Mitf null allele, and it induces such a transdifferentiation in embryos that are either heterozygous for the Mitf null allele or homozygous for an RPE-specific hypomorphic Mitf allele generated by targeted mutation. Conversely, an increase in Pax6 gene dose interferes with transdifferentiation even in homozygous Mitf null embryos. Gene expression analyses show that, together with MITF or its paralog TFEC, PAX6 suppresses the expression of Fgf15 and Dkk3. Explant culture experiments indicate that a combination of FGF and DKK3 promote retina formation by inhibiting canonical WNT signaling and stimulating the expression of retinogenic genes, including Six6 and Vsx2. Our results demonstrate that in conjunction with Mitf/Tfec Pax6 acts as an anti-retinogenic factor, whereas in conjunction with retinogenic genes it acts as a pro-retinogenic factor. The results suggest that careful manipulation of the Pax6 regulatory circuit may facilitate the generation of retinal and pigment epithelium cells from embryonic or induced pluripotent stem cells.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pgen.1002757

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 43

  1. [1]Bharti K Nguyen MT Skuntz S Bertuzzi S Arnheiter H 2006 The other pigment cell: specification and development of the pigmented epithelium of the vertebrate eye. Pigment Cell Research 19 380 394 16965267 PMC1564434
  2. [2]Karl MO Reh TA 2010 Regenerative medicine for retinal diseases: activating endogenous repair mechanisms. Trends Mol Med 16 193 202 20303826 PMC2854262
  3. [3]Miller SS Maminishkis A Li R Adijanto J 2010 Retinal Pigment Epithelium: Cytokine Modulation of Epithelial Physiology. Dartt DA Encyclopedia of the Eye: Oxford University Academic Press 89 100 editor
  4. [4]Hemesath TJ Steingrimsson E McGill G Hansen MJ Vaught J 1994 microphthalmia, a critical factor in melanocyte development, defines a discrete transcription factor family. Genes Dev 8 2770 2780 7958932
  5. [5]Hodgkinson CA Moore KJ Nakayama A Steingrimsson E Copeland NG 1993 Mutations at the mouse microphthalmia locus are associated with defects in a gene encoding a novel basic-helix-loop-helix-zipper protein. Cell 74 395 404 8343963
  6. [6]Horsford DJ Nguyen MT Sellar GC Kothary R Arnheiter H 2005 Chx10 repression of Mitf is required for the maintenance of mammalian neuroretinal identity. Development 132 177 187 15576400
  7. [7]Nguyen M Arnheiter H 2000 Signaling and transcriptional regulation in early mammalian eye development: a link between FGF and MITF. Development 127 3581 3591 10903182
  8. [8]Fujimura N Taketo MM Mori M Korinek V Kozmik Z 2009 Spatial and temporal regulation of Wnt/beta-catenin signaling is essential for development of the retinal pigment epithelium. Dev Biol 334 31 45 19596317
  9. [9]Grocott T Johnson S Bailey AP Streit A 2011 Neural crest cells organize the eye via TGF-beta and canonical Wnt signalling. Nat Commun 2 265 21468017 PMC3104559
  10. [10]Müller F Rohrer H Vogel-Hopker A 2007 Bone morphogenetic proteins specify the retinal pigment epithelium in the chick embryo. Development 134 3483 3493 17728349
  11. [11]Westenskow P Piccolo S Fuhrmann S 2009 Beta-catenin controls differentiation of the retinal pigment epithelium in the mouse optic cup by regulating Mitf and Otx2 expression. Development 136 2505 2510 19553286 PMC2709060
  12. [12]Fuhrmann S Levine EM Reh TA 2000 Extraocular mesenchyme patterns the optic vesicle during early eye development in the embryonic chick. Development 127 4599 4609 11023863
  13. [13]Bumsted KM Barnstable CJ 2000 Dorsal retinal pigment epithelium differentiates as neural retina in the microphthalmia (mi/mi) mouse. Invest Ophthalmol Vis Sci 41 903 908 10711712
  14. [14]Bharti K Liu W Csermely T Bertuzzi S Arnheiter H 2008 Alternative promoter use in eye development: the complex role and regulation of the transcription factor MITF. Development 135 1169 1178 18272592 PMC2276638
  15. [15]Grindley JC Davidson DR Hill RE 1995 The role of Pax-6 in eye and nasal development. Development 121 1433 1442 7789273
  16. [16]Martinez-Morales JR Rodrigo I Bovolenta P 2004 Eye development: a view from the retina pigmented epithelium. Bioessays 26 766 777 15221858
  17. [17]Quinn JC West JD Hill RE 1996 Multiple functions for Pax6 in mouse eye and nasal development. Genes Dev 10 435 446 8600027
  18. [18]Hill RE Favor J Hogan BL Ton CC Saunders GF 1991 Mouse small eye results from mutations in a paired-like homeobox-containing gene. Nature 354 522 525 1684639
  19. [19]Marquardt T Ashery-Padan R Andrejewski N Scardigli R Guillemot F 2001 Pax6 is required for the multipotent state of retinal progenitor cells. Cell 105 43 55 11301001
  20. [20]Schedl A Ross A Lee M Engelkamp D Rashbass P 1996 Influence of PAX6 gene dosage on development: overexpression causes severe eye abnormalities. Cell 86 71 82 8689689
  21. [21]Manuel M Pratt T Liu M Jeffery G Price DJ 2008 Overexpression of Pax6 results in microphthalmia, retinal dysplasia and defective retinal ganglion cell axon guidance. BMC Dev Biol 8 59 18507827 PMC2422841
  22. [22]Collinson JM Quinn JC Hill RE West JD 2003 The roles of Pax6 in the cornea, retina, and olfactory epithelium of the developing mouse embryo. Dev Biol 255 303 312 12648492
  23. [23]Planque N Leconte L Coquelle FM Martin P Saule S 2001 Specific Pax-6/microphthalmia transcription factor interactions involve their DNA-binding domains and inhibit transcriptional properties of both proteins. J Biol Chem 276 29330 29337 11350962
  24. [24]Bäumer N Marquardt T Stoykova A Spieler D Treichel D 2003 Retinal pigmented epithelium determination requires the redundant activities of Pax2 and Pax6. Development 130 2903 2915 12756174
  25. [25]Koso H Iida A Tabata Y Baba Y Satoh S 2008 CD138/syndecan-1 and SSEA-1 mark distinct populations of developing ciliary epithelium that are regulated differentially by Wnt signal. Stem Cells 26 3162 3171 18787210
  26. [26]Mui SH Kim JW Lemke G Bertuzzi S 2005 Vax genes ventralize the embryonic eye. Genes Dev 19 1249 1259 15905411 PMC1132010
  27. [27]Tang K Xie X Park JI Jamrich M Tsai S 2010 COUP-TFs regulate eye development by controlling factors essential for optic vesicle morphogenesis. Development 137 725 734 20147377 PMC2827684
  28. [28]Rowan S Chen CM Young TL Fisher DE Cepko CL 2004 Transdifferentiation of the retina into pigmented cells in ocular retardation mice defines a new function of the homeodomain gene Chx10. Development 131 5139 5152 15459106
  29. [29]Murisier F Guichard S Beermann F 2007 Distinct distal regulatory elements control tyrosinase expression in melanocytes and the retinal pigment epithelium. Dev Biol 303 838 847 17196956
  30. [30]Kuiper RP Schepens M Thijssen J Schoenmakers EF van Kessel AG 2004 Regulation of the MiTF/TFE bHLH-LZ transcription factors through restricted spatial expression and alternative splicing of functional domains. Nucleic Acids Res 32 2315 2322 15118077 PMC419459
  31. [31]Guillemot F Cepko CL 1992 Retinal fate and ganglion cell differentiation are potentiated by acidic FGF in an in vitro assay of early retinal development. Development 114 743 754 1377624
  32. [32]Zhao S Thornquist SC Barnstable CJ 1995 In vitro transdifferentiation of embryonic rat retinal pigment epithelium to neural retina. Brain Res 677 300 310 7552256
  33. [33]Niehrs C 2006 Function and biological roles of the Dickkopf family of Wnt modulators. Oncogene 25 7469 7481 17143291
  34. [34]Yue W Sun Q Dacic S Landreneau RJ Siegfried JM 2008 Downregulation of Dkk3 activates beta-catenin/TCF-4 signaling in lung cancer. Carcinogenesis 29 84 92 18048388
  35. [35]Nakamura RE Hunter DD Yi H Brunken WJ Hackam AS 2007 Identification of two novel activities of the Wnt signaling regulator Dickkopf 3 and characterization of its expression in the mouse retina. BMC Cell Biol 8 52 18093317 PMC2233618
  36. [36]Guillemot F Zimmer C 2011 From cradle to grave: The multiple roles of fibroblast growth factors in neural development. Neuron 71 574 588 21867876
  37. [37]Mohamed OA Clarke HJ Dufort D 2004 Beta-catenin signaling marks the prospective site of primitive streak formation in the mouse embryo. Dev Dyn 231 416 424 15366019
  38. [38]Zuber ME Gestri G Viczian AS Barsacchi G Harris WA 2003 Specification of the vertebrate eye by a network of eye field transcription factors. Development 130 5155 5167 12944429
  39. [39]Eiraku M Takata N Ishibashi H Kawada M Sakakura E 2011 Self-organizing optic-cup morphogenesis in three-dimensional culture. Nature 472 51 56 21475194
  40. [40]Matsushima D Heavner W Pevny LH 2011 Combinatorial regulation of optic cup progenitor cell fate by SOX2 and PAX6. Development 138 443 454 21205789 PMC3014633
  41. [41]Hyer J Mima T Mikawa T 1998 FGF1 patterns the optic vesicle by directing the placement of the neural retina domain. Development 125 869 877 9449669
  42. [42]Bharti K Miller SS Arnheiter H 2011 The new paradigm: retinal pigment epithelium cells generated from embryonic or induced pluripotent stem cells. Pigment Cell Melanoma Res 24 21 34 20846177 PMC3021640
  43. [43]Campos M Amaral J Becerra SP Fariss RN 2006 A novel imaging technique for experimental choroidal neovascularization. Invest Ophthalmol Vis Sci 47 5163 5170 17122098

Cited by

Sorted by citations

Authors
Bharti, K., Gasper, M., Ou, J., Brucato, M., Clore-Gronenborn, K., Pickel, J., & Arnheiter, H.
Affiliations
National Institutes of Health (US)
Corresponding author
Bharti, Kapil
ORCID
Venue
PLoS Genetics
Publisher
Public Library of Science (PLoS)
Published
2012-07-05
Volume
8
Issue
7
Pages
e1002757
ISSN
  • 1553-7390 (ISSN-L)
  • 1553-7404
DOI
10.1371/journal.pgen.1002757
PMC
PMC3390378
OpenAlex
W2035797804
Cited by
158
References
0
Citations by year
2026: 7 · 2025: 5 · 2024: 8 · 2023: 11 · 2022: 12 · 2021: 19 · 2020: 16 · 2019: 13 · 2018: 12 · 2017: 7 · 2016: 9 · 2015: 8 · 2014: 17 · 2013: 13 · 2012: 1
FWCI
4.69
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS Genetics (publishedVersion, public-domain)
  • Europe PMC (PubMed Central) (publishedVersion, public-domain)
  • PubMed Central (publishedVersion, public-domain)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
Funders
  • National Institutes of Health
  • National Human Genome Research Institute
  • National Institute of Neurological Disorders and Stroke
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Ocular Disorders and Treatments
Keywords
Pax6, MITF, TFEC, Wnt signaling, FGF15, DKK3, SIX6, VSX2, eye development, gene dosage, induced pluripotent stem cells
MeSH terms
PAX6 Transcription Factor, Animals, Eye Proteins, Fibroblast Growth Factors, Repressor Proteins, Retina, Homeodomain Proteins, Gene Expression Regulation, Developmental, Gene Dosage, Intercellular Signaling Peptides and Proteins, Embryonic Development, Adaptor Proteins, Signal Transducing, Mice, Microphthalmia-Associated Transcription Factor, Paired Box Transcription Factors, Cell Transdifferentiation, Retinal Pigment Epithelium, Wnt Signaling Pathway

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review