Neurospine.  2018 Mar;15(1):42-53. 10.14245/ns.1836010.005.

The Effect of Recombinant Tyrosine Hydroxylase Expression on the Neurogenic Differentiation Potency of Mesenchymal Stem Cells

• Turac G
• Duruksu G
• Karaoz E

• KMID: 2413271
• DOI: http://doi.org/10.14245/ns.1836010.005

Abstract

OBJECTIVE
Tyrosine hydroxylase (TH) is a rate-limiting enzyme in dopamine synthesis, making the enhancement of its activity a target for ensuring sufficient dopamine levels. Rat bone marrow mesenchymal stem cells (rBM-MSCs) are known to synthesize TH after differentiating into neuronal cells through chemical induction, but the effect of its ectopic expression on these cells has not yet been determined. This study investigated the effects of ectopic recombinant TH expression on the stemness characteristics of rBM-MSCs.
METHODS
After cloning, a cell line with stable TH expression was maintained, and the proliferation, the gene expression profile, and differentiation potential of rBM-MSCs were analyzed. Analysis of the cells showed an increment in the proliferation rate that could be reversed by the neutralization of TH.
RESULTS
The constitutive expression of TH in rBM-MSCs was successfully implemented, without significantly affecting their osteogenic and adipogenic differentiation potential. TH expression improved the expression of other neuronal markers, such as glial fibrillary acidic protein, β-tubulin, nestin, and c-Fos, confirming the neurogenic differentiation capacity of the stem cells. The expression of brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) significantly increased after the chemical induction of neurogenic differentiation.
CONCLUSION
In this study, the expression of recombinant TH improved the neuroprotective effect of MSCs by upregulating the expression of BDNF and CNTF. Although the neuronal markers were upregulated, the expression of recombinant TH alone in rBM-MSCs was not sufficient for MSCs to differentiate into neurogenic cell lines.

Keyword

Tyrosine hydroxylase; Differentiation; Gene expression; Neuroprotection; Mesenchymal stem cell

MeSH Terms

Animals
Bone Marrow
Brain-Derived Neurotrophic Factor
Cell Line
Ciliary Neurotrophic Factor
Clone Cells
Cloning, Organism
Dopamine
Ectopic Gene Expression
Gene Expression
Glial Fibrillary Acidic Protein
Mesenchymal Stromal Cells*
Nestin
Neurons
Neuroprotection
Neuroprotective Agents
Rats
Stem Cells
Transcriptome
Tyrosine 3-Monooxygenase*
Tyrosine*
Brain-Derived Neurotrophic Factor
Ciliary Neurotrophic Factor
Dopamine
Glial Fibrillary Acidic Protein
Neuroprotective Agents
Tyrosine
Tyrosine 3-Monooxygenase