Skip to main content
SpringerLink
Log in

Atoh7 promotes retinal Müller cell differentiation into retinal ganglion cells

  • Original Research
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

Glaucoma is one of the leading eye diseases due to the death of retinal ganglion cells. Increasing evidence suggests that retinal Müller cells exhibit the characteristics of retinal progenitor cells and can differentiate to neurons in injured retinas under certain conditions. However, the number of ganglion cells differentiated from retinal Müller cells falls far short of therapeutic needs. This study aimed to promote the differentiation of retinal Müller cells into ganglion cells by introducing Atoh7 into the stem cells dedifferentiated from retinal Müller cells. Rat retinal Müller cells were isolated and dedifferentiated into stem cells, which were transfected with PEGFP-N1 or PEGFP-N1-Atoh7 vector, and then further induced to differentiate into ganglion cells. The proportion of ganglion cells differentiated from Atoh7-tranfected stem cells was significantly higher than that of control transfected or untransfected cells. In summary, Atoh7 promotes the differentiation of retinal Müller cells into retinal ganglion cells. This may open a new avenue for gene therapy of glaucoma by promoting optic nerve regeneration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Aberg MA, Ryttsén F, Hellgren G, Lindell K, Rosengren LE, MacLennan AJ, Carlsson B, Orwar O, Eriksson PS (2001) Selective introduction of antisense oligonucleotides into single adult CNS progenitor cells using electroporation demonstrates the requirement of STAT3 activation for CNTF-induced gliogenesis. Mol Cell Neurosci 17:426–443

    Article  CAS  Google Scholar 

  • Abrahan CE, Insua MF, Politi LE, German OL, Rotstein NP (2009) Oxidative stress promotes proliferation and dedifferentiation of retina glial cells in vitro. J Neurosci Res 87:964–977

    Article  CAS  Google Scholar 

  • Ahmed S, Reynolds BA, Weiss S (1995) BDNF enhances the differentiation but not the survival of CNS stem cell-derived neuronal precursors. J Neurosci 15:5765–5778

    CAS  Google Scholar 

  • Baum C, Forster P, Hegewisch-Becker S, Harbers K (1994) An optimized electroporation protocol applicable to a wide range of cell lines. Biotechniques 17:1058–1062

    CAS  Google Scholar 

  • Bringmann A, Pannicke T, Grosche J, Francke M, Wiedemann P, Skatchkov SN, Osborne NN, Reichenbach A (2006) Müller cells in the healthy and diseased retina. Prog Retin Eye Res 25:397–424

    Article  CAS  Google Scholar 

  • Brown NL, Kanekar S, Vetter ML, Tucker PK, Gemza DL, Glaser T (1998) Atoh7 encodes a murine basic helix-loop-helix transcription factor expressed during early stages of retinal neurogenesis. Development 125:4821–4833

    CAS  Google Scholar 

  • Brown NL, Patel S, Brzezinski J, Glaser T (2001) Atoh7 is required for retinal ganglion cell and optic nerve formation. Development 128:2497–2508

    CAS  Google Scholar 

  • Cepko CL (1999) The roles of intrinstic and extrinsic cues and bHLH genes in the determination of retinal cell fates. Curr Opin Neurobiol 9:37–46

    Article  CAS  Google Scholar 

  • Cesnulevicius K, Timmer M, Wesemann M, Thomas T, Barkhausen T, Grothe C (2006) Nucleofection is the most efficient nonviral transfection method for neuronal stem cells derived from ventral mesencephali with no changes in cell composition or dopaminergic fate. Stem Cells 24:2776–2791

    Article  CAS  Google Scholar 

  • Cho JH, Mao CA, Klein WH (2012) Adult mice transplanted with embryonic retinal progenitor cells: new approach for repairing damaged optic nerves. Mol Vis 18:2658–2672

    CAS  Google Scholar 

  • Das AV, Mallya KB, Zhao X, Ahmad F, Bhattacharya S, Thoreson WB, Hegde GV, Ahmad I (2006) Neural stem cell properties of Müller glia in the mammalian retina: regulation by Notch and Wnt signaling. Dev Biol 299:283–302

    Article  CAS  Google Scholar 

  • Fimbel SM, Montgomery JE, Burket CT, Hyde DR (2007) Regeneration of inner retinal neurons after intravitreal injection of ouabain in zebrafish. J Neurosci 27:1712–1724

    Article  CAS  Google Scholar 

  • Geoffroy CG, Raineteau O (2007) A cre-lox approach for transient transgene expression in neural precursor cells and long-term tracking of their progeny in vitro and in vivo. BMC Dev Biol 7:45

    Article  Google Scholar 

  • Golzio M, Mora MP, Raynaud C, Delteil C, Teissié J, Rols MP (1998) Control by osmotic pressure of voltage-induced permeabilization and gene transfer in mammalian cells. Biophys J 74:3015–3022

    Article  CAS  Google Scholar 

  • Guan K, Chang H, Rolletschek A, Wobus AM (2001) Embryonic stem cell-derived neurogenesis: retinoic acid induction and lineage selection of neuronal cells. Cell Tissue Res 305:171–176

    Article  CAS  Google Scholar 

  • Helledie T, Nurcombe V, Cool SM (2008) A simple and reliable electroporation method for human bone marrow mesenchymal stem cells. Stem Cells Del 17:837–848

    Article  CAS  Google Scholar 

  • Kim YC, Shim JW, Oh YJ, Son H, Lee YS, Lee SH (2002) Co-transfection with cDNA encoding the Bcl family of anti-apoptotic proteins improves the efficiency of transfection in primary fetal neural stem cells. J Neurosci Methods 117:153–158

    Article  CAS  Google Scholar 

  • Kubota A, Nishida K, Nakashima K, Tano Y (2006) Conversion of mammalian Müller glia cells into a neuronal lineage by in vitro aggregate. Biochem Biophys Res Commun 351:514–520

    Article  CAS  Google Scholar 

  • Lawrence JM, Singhal S, Bhatia B, Keegan DJ, Reh TA, Luthert PJ, Khaw PT, Limb GA (2007) MIO-M1 cells and similar muller glial cell lines derived from adult human retina exhibit neural stem cell characteristics. Stem Cells 25:2033–2043

    Article  CAS  Google Scholar 

  • Ooto S, Akagi T, Kageyama R, Akita J, Mandai M, Honda Y, Takahashi M (2004) Potential for neural regeneration after neurotoxic injury in the adult mammalian retina. Proc Natl Acad Sci USA 101:13654–13659

    Article  CAS  Google Scholar 

  • Sarthy VP, Brodjian SJ, Dutt K, Kennedy BN, French RP, Crabb JW (1998) Establishment and characterization of a retinal Müller cell line. Invest Ophthalmol Vis Sci 39:212–216

    CAS  Google Scholar 

  • Tropepe V, Coles BL, Chiasson BJ, Horsford DJ, Elia AJ, McInnes RR, van der Kooy D, Tropepe V, Coles BL, Chiasson BJ (2000) Retinal stem cells in the adult mammalian eye. Science 287:2032–2036

    Article  CAS  Google Scholar 

  • Wang SW, Kim BS, Ding K, Wang H, Sun D, Johnson RL, Klein WH, Gan L (2001) Requirement for Atoh7 in the development of retinal ganglion cells. Genes Dev 15:24–29

    Article  CAS  Google Scholar 

  • Yang Z, Ding K, Pan L, Deng M, Gan L (2003) Atoh7 determines the competence state of retinal ganglion cell progenitors. Dev Biol 264:240–254

    Article  CAS  Google Scholar 

  • Yao J, Sun X, Wang Y, Xu G, Qian J (2007) Atoh7 promotes retinal ganglion cell expression patterns in retinal progenitor cells. Mol Vis 13:1066–1072

    CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by Grant from National Scientific Foundation of China (No. NSFC 81170844) and State Key Lab of Medical Genetics of China (No. 1989DA105084). We are grateful to Prof. Nadean L. Brown (University of Clicinnati College of Medicine) for kindly gifting the PBSIISK-Atoh7 plasmid. We thank Dr. Jingwei Chi, for her valuable advice and guidance; Dr. Yaping Yan, Dr. Xiaoyun Mo and Dr. Jieqiong Tan, for their helpful suggestion; Dr. Shanshan Zhang and Chunjiang Liu for technical assistance.

Author information

Authors and Affiliations

  1. Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, 410008, China

    Wei-tao Song, Qi Zeng & Xiao-bo Xia

  2. State Key Lab of Medical Genetics of China, Central South University, Changsha, China

    Kun Xia & Qian Pan

Authors
  1. Wei-tao Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qi Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiao-bo Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kun Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qian Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xiao-bo Xia.

Additional information

Wei-tao Song and Qi Zeng contributed equally to this work

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Song, Wt., Zeng, Q., Xia, Xb. et al. Atoh7 promotes retinal Müller cell differentiation into retinal ganglion cells. Cytotechnology 68, 267–277 (2016). https://doi.org/10.1007/s10616-014-9777-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-014-9777-1

Keywords

Search

Navigation