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Applied Biochemistry and Biotechnology

, Volume 177, Issue 1, pp 48–62 | Cite as

Overexpressed Calponin3 by Subsonic Vibration Induces Neural Differentiation of hUC-MSCs by Regulating the Ionotropic Glutamate Receptor

  • Hyun-Jung Kim
  • Jin-Hee Kim
  • Yeo-Ju Song
  • Young-Kwon Seo
  • Jung-Keug Park
  • Chan-Wha KimEmail author
Article

Abstract

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.

Keywords

Subsonic vibration Mesenchymal stem cells Neural differentiation Proteomics CNN3 Glutamate receptor 

Abbreviations

hUC-MSC

Human umbilical cords derived mesenchymal stem cell

SV

Sonic vibration

2-DE

Two-dimensional electrophoresis

LC-ESI MS/MS

Liquid chromatography electrospray ionization tandem mass spectrometry

CNN3

Calponnin3

PSD95

Postsynaptic density protein 95

GluR1

Glutamate receptor 1

AMPA

2-Amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid

NR1

N-Methyl-d-aspartate

PBS

Phosphate-buffered saline

IEF

Isoelectric focusing

IAA

Iodoacetamide

BPB

Bromophenol blue

SEM

Standard error of the mean

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

BSA

Bovine serum albumin

FBS

Fetal bovine serum

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide electrophoresis

TBST

Tween-20

Notes

Acknowledgments

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.

Author Contributions

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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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Hyun-Jung Kim
    • 1
  • Jin-Hee Kim
    • 1
  • Yeo-Ju Song
    • 1
  • Young-Kwon Seo
    • 2
  • Jung-Keug Park
    • 2
  • Chan-Wha Kim
    • 1
    Email author
  1. 1.College of Life of Life Sciences and BiotechnologyKorea UniversitySeoulKorea
  2. 2.Department of Medical BiotechnologyDongguk UniversitySeoulKorea

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