April 2010
Volume 51, Issue 13
ARVO Annual Meeting Abstract  |   April 2010
Characterization of Retinal Stem Cells and Their Math5-Induced Differentiation by Lentivirus-Mediated Gene Transfer
Author Affiliations & Notes
  • X. Zhang
    Neuro-Ophthalmology, Xuwu Hospital, Beijing, China
  • Footnotes
    Commercial Relationships  X. Zhang, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 729. doi:
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      X. Zhang; Characterization of Retinal Stem Cells and Their Math5-Induced Differentiation by Lentivirus-Mediated Gene Transfer. Invest. Ophthalmol. Vis. Sci. 2010;51(13):729.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: : math5

Methods: : RSCs were isolated from neural retina of postnatal day 10 mice and grown as neurospheres in a defined serum-free culture medium. math5 coding sequence was subcloned into feline immunodeficiency virus (FIV) vector with N-terminal c-myc tag. RSCs were infected with FIV expressing c-myc-tagged math5 or control c-myc FIV. The expression of math5, undifferentiated progenitor-specific marker Nestin and neuron-specific DNA-binding protein NeuN (Neuronal Nuclei) were characterized in RSCs by immunocytochemistry using marker-specific antibodies.

Results: : c-myc expression was detected by immunocytochemistry in FIV-infected RSCs, suggesting that RSCs were successfully infected by FIV with the expression of c-myc-math5. Immunocytochemical analysis revealed that both FIV-math5-infected and mock infected RSCs expressed stem cell marker Nestin. However, NeuN, a neuron-specific marker, was expressed only in the RSCs infected with math5, but not in mock infected RSCs.

Conclusions: : These results demonstrate that expression of math5 induces NeuN expression in RSCs, suggesting that math5 promotes RSC differentiation toward RGCs by upregulating neuron-specific gene expression. This study also reveals that virus-mediated gene transfer can facilitate the investigation of gene-specific regulation of RSC differentiation.

Keywords: gene transfer/gene therapy • differentiation • gene/expression 

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