Overexpressed Calponin3 by Subsonic Vibration Induces Neural Differentiation of hUC-MSCs by Regulating the Ionotropic Glutamate Receptor
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In this study, we used proteomics to investigate the effects of sonic vibration (SV) on mesenchymal stem cells derived from human umbilical cords (hUC-MSCs) during neural differentiation to understand how SV enhances neural differentiation of hUC-MSCs. We investigated the levels of gene and protein related to neural differentiation after 3 or 5 days in a group treated with 40-Hz SV. In addition, protein expression patterns were compared between the control and the 40-Hz SV-treated hUC-MSC groups via a proteomic approach. Among these proteins, calponin3 (CNN3) was confirmed to have 299 % higher expression in the 40-Hz SV stimulated hUC-MSCs group than that in the control by Western blotting. Notably, overexpression of CNN3-GFP in Chinese hamster ovary (CHO)-K1 cells had positive effects on the stability and reorganization of F-actin compared with that in GFP-transfected cells. Moreover, CNN3 changed the morphology of the cells by making a neurite-like form. After being subjected to SV, messenger RNA (mRNA) levels of glutamate receptors such as PSD95, GluR1, and NR1 as well as intracellular calcium levels were upregulated. These results suggest that the activity of glutamate receptors increased because of CNN3 characteristics. Taken together, these results demonstrate that overexpressed CNN3 during SV increases expression of glutamate receptors and promotes functional neural differentiation of hUC-MSCs.
KeywordsSubsonic vibration Mesenchymal stem cells Neural differentiation Proteomics CNN3 Glutamate receptor
Human umbilical cords derived mesenchymal stem cell
- LC-ESI MS/MS
Liquid chromatography electrospray ionization tandem mass spectrometry
Postsynaptic density protein 95
Glutamate receptor 1
Standard error of the mean
Bovine serum albumin
Fetal bovine serum
Sodium dodecyl sulfate polyacrylamide electrophoresis
This study was supported by a Korea University Grant and the Pioneer Research Program of the National Research Foundation of Korea, founded by the Ministry of Education, Science, and Technology (2009-0082946).
Conflict of Interest
The authors declare no conflict of interest and ethical approval.
HJ Kim conducted cell culture, BrdU proliferation assay, RT-PCR immunofluorescence staining, Western blotting, and manuscript writing. JH Kim conducted transfection of CNN3 and immunofluorescence staining, YJ Song performed two-dimensional electrophoresis and ESI-Q-TOF MS/MS, and YK Seo conducted intracellular calcium level. JK Park and CW Kim were involved in the experimental design of the study and in the interpretation of the data and manuscript editing. All the authors declare that they have no competing interests.
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