Abstract
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