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Neuronal differentiation of immortalized neural stem cells by overexpression of neurogenin1

Neuronal differentiation of immortalized neural stem cells by overexpression of neurogenin1

Society for Neuroscience Abstracts 27(1): 1233

The basic helix-loop-helix transcription factor neurogenin1 (ngn1) has been shown to play important roles in mammalian neurogenesis. In this study, we investigated the neurogenic potential of ngn1 in neuroblastoma and neural stem cells. To induce or block the expression of ngn1, the expression vectors for the full length and C-terminal truncated mutant of ngn1 were generated. Transient expression of the full-length ngn1 and its dimerization partner, E47, synergically increased luciferase activity of the reporter gene about 40-fold. In contrast, expression of the C-terminal truncated mutant of ngn1 partially repressed the reporter gene that was induced by the full-length ngn1. The neurogenic effect of ngn1 was verified using F11 neuroblastoma cells that could be induced to differentiate into neurons in the presence of cAMP. Overexpression of the full-length ngn1 induced neurite outgrowth in F11 cells in the absence of cAMP. In contrast, the C-terminal truncated mutant of ngn1 did not alter neurite outgrowth induced by cAMP in F11 cells. In human neural stem cells that were generated by immortalizing human fetal neural stem cells with v-myc, stable expression of ngn1 induced expression of a neuron specific marker, NCAM but not an astrocyte-specific marker, GFAP. After transplanted into neonatal mouse brain, the ngn1-expressing stable cells differentiated into neurons but not to astrocytes. These results suggest that ngn1 may be a useful tool to derive undetermined stem cells into a neuronal fate.

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Accession: 035380729

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