In vivo bioluminescence imaging for newly identified neurogenic microRNAs related to neuronal differentiation induced by neurogenin 1 (Ngn 1) via microarray analysis

Objectives MicroRNAs (miRNAs) are known as critical gene regulator and are involved in regulation of neuronal differentiation. The aim of this study is to identify neurogenic miRNA induced by Ngn1 during neurogenesis via miRNA microarray.

Methods MiRNA microarray was performed from isolated total RNA using agilent microarray scanner. Functional annotation for target genes of miRNAs was searched by DAVID v6.7 database. The 3′-UTR-based luciferase system containing 3 copy binding sequence for miRNA was used to observe expression of identified miRNA.

Results Microarray results in Ngn1-overproduced F11 cells showed 240 upregulated miRNAs including miR-132 were considered as neurogenic miRNAs. We found that target genes of miRNAs were associated with neurogenesis, axon guidance and cell cycle. F11 cells generated neurite outgrowth within 3 days after the newly identified miRNA treatment. Immunofluorescence staining represented that βIII-tubulin, NeuroD and MAP2 neuronal marker expression was seen 3 days after treatment of identified miRNA. An in vivo study showed that bioluminescence signals were gradually decreased in identified miRNA treated group until 2 days.

Conclusions In this study, we analyzed the miRNA expression profiling in Ngn1-treated F11 cells, via miRNA microarray. This study demonstrates that newly identified neurogenic microRNAs could be used as new potential inducer for neuronal differentiation in terms of stem cell-based therapy.