Purpose To directly convert mouse fibroblast to functional neurons by adenovirus carrying neural specific transcription factors.
Methods We infected mouse embryonic and adult fibroblasts with adenoviruses carrying transcription factors ABM (Ascl1, Brn2, and Myt1l) or ABN (Ascl1, Brn2, and Ngn2). We examined the properties of induced neurons (iNs) by immunostaining of multiple cell markers and electrophysiological techniques. Quantitative PCR and southern blot were also used to test whether adenovirus mediated reprogramming caused virus vectors integration into host genome.
Results Both combinations of ABM and ABN were sufficient to convert fibroblasts to neurons, with a relative higher efficiency in ABN combination group. iNs expressed neural specific markers of Tuj1, MAP2a, NeuN, Synapsin, as well as excitatory neuron marker vGlut, and inhibitory neuron marker GABA. iNs also exhibit action potential. Quantitative PCR and southern blot showed that adenovirus mediated reprogramming didn’t cause virus vectors integration into host genome.
Conclusions Adenoviral transduction may be used as a safer tool for the conversion of fibroblasts to neurons without viral integration risk. And our study shed new light on the therapy of neural degeneration diseases, including retinal degeneration diseases.
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