Adenovirus-mediated RAR-β over-expression enhances ATRA-induced neuronal differentiation of rat mesenchymal stem cells

4Citations
Citations of this article
11Readers
Mendeley users who have this article in their library.

Abstract

Introduction: The retinoic acid (RA) signaling pathway plays important roles in neural development. All-trans retinoic acid (ATRA) activates the RA signal by regulating RAR-β in mesenchymal stem cell (MSC)-derived neuron cells. Here, we try to investigate whether RAR-β over-expression can affect neuronal differentiation of MSCs. Material and methods: The RAR-β gene was constructed into adenovirus Ad-RAR-β by using the AdEasy system. The MSCs were infected with Ad-RAR-β. Real timepolymerase chain reaction (RT-PCR), Western blot and immunofluorescence were performed to detect the expression and localization of RAR-β. The MSCs were treated with 1 μmol/l ATRA and modified neuronal induction medium (MNM). Soma size and axon length of induced neurons were measured. Neural specific markers were detected by RT-PCR, western blot and immunofluorescence to evaluate neuronal differentiation. Results: The 1300 bp fragment of RAR-β gene was confirmed to be correctly cloned in the adenovirus vector. Cloudiness amplification of Ad-RAR- β was observed in HEK293 cells during package. After 48 h of Ad-RAR-β infection, about 70% of MSCs were RFP-positive. RAR-β expression was increased by about 1988-fold and located in the nucleus. RAR-β over-expression did not affect neuronal differentiation efficiency; however, soma size of induced neuron cells enlarged from 716.25 ±95.96 μm2 to 1160.12 ±352.65 μm 2 and axon length from 64.17 ±11.88 μm to 83.98 ±13.69 μm. Neural markers other than nestin - NSE, MAP-2, Tau, and Tuj1 - were increased by 4- to 11-fold in RAR- β over-expressed neuron cells with ATRA/MNM induction compared with the Ad-null control group. Conclusions: Our results have demonstrated that adenovirus-mediated RAR-β over-expression could facilitate neuron cell types of MSCs in vitro, indicating that the RAR-β-activated RA signal might be a vital factor in neuronal differentiation. Copyright © 2013 Termedia & Banach.

Cite

CITATION STYLE

APA

Bi, Y., Gong, M., He, Y., Wei, X., Chen, J., & Li, T. (2013). Adenovirus-mediated RAR-β over-expression enhances ATRA-induced neuronal differentiation of rat mesenchymal stem cells. Archives of Medical Science, 9(2), 314–322. https://doi.org/10.5114/aoms.2012.31410

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free