Effect of NTN and Lmx1α on the Notch Signaling Pathway during the Differentiation of Human Bone Marrow Mesenchymal Stem Cells into Dopaminergic Neuron-Like Cells.

Shuaiyao Lu,Giuseppina Martella,Xuantao Yang,Juanjuan Zhang,Bo Yang,Wanpu Wang,Lilin Luo,Jinhua Zhang,Yuxin Xie,Linhui Li

doi:10.1155/2021/6676709

Abstract

Human bone marrow mesenchymal stem cells (h-BMSCs) have the potential to differentiate into dopaminergic neuron-like cells to treat Parkinson's disease. The Notch signaling pathway has been implicated in the regulation of cell fate decisions such as differentiation of BMSCs. This study investigated changes in the expression of Notch-related genes in the differentiation of BMSCs in vitro into dopaminergic (DA) neuron-like cells. BMSCs transfected with empty lentiviral vectors served as the control group and those transfected with NTN and Lmx1α recombinant lentiviral vectors served as the experimental group. After induction and culture of NTN and Lmx1α-transfected h-BMSCs for 21 days, the cells exhibited features of dopaminergic neuron-like cells, which were observed by transmission and scanning electron microscopy and verified by immunofluorescence of tyrosine hydroxylase (TH) and dopamine transporter (DAT). These induced cells could secrete dopamine and had basic action potentials. Expression of the neural stem cell (NSC) markers, including octamer-binding protein (Oct4), paired box gene 6 (Pax6), and sex determining region Y-box 1 (SOX1), increased on day 14 of induction and decreased on day 21 of induction during differentiation. The human Notch signaling pathway PCR array showed a differential expression of Notch-related genes during the differentiation of h-BMSCs into DA neuron-like cells in vitro relative to that in the control group. In conclusion, h-BMSCs overexpressing NTN and Lmx1α can successfully be induced to differentiate into dopaminergic neuron-like cells with a neuronal phenotype exhibiting fundamental biological functions in vitro, and NTN and Lmx1α may affect the expression of Notch-related genes during differentiation.

Highlights

Parkinson’s disease (PD) is an age-related neurodegenerative disease that is clinically characterized by the death of dopaminergic (DA) neurons and the formation of Lewy bodies [1, 2]. e mainstay of PD management is drug therapy, but drug therapy cannot prevent disease development
We demonstrated that DA can be efficiently induced from bone marrow mesenchymal stem cells (BMSCs) overexpressing the LIM homeobox transcription factor 1α (Lmx1α) and neurturin (NTN) [4]
Grayscale analysis, and PCR demonstrated that the NTN and Lmx1α expression levels significantly differed between the transfection and control groups (p < 0.005; Figures 1(b)–1(d)). ese results confirm the successful transfection and efficient expression of the NTN and Lmx1α in Human bone marrow mesenchymal stem cells (h-BMSCs)

Summary

Introduction

Parkinson’s disease (PD) is an age-related neurodegenerative disease that is clinically characterized by the death of dopaminergic (DA) neurons and the formation of Lewy bodies [1, 2]. e mainstay of PD management is drug therapy, but drug therapy cannot prevent disease development. We demonstrated that DA can be efficiently induced from BMSCs overexpressing the LIM homeobox transcription factor 1α (Lmx1α) and neurturin (NTN) [4]. When these BMSCs were grafted into 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydro pyridine(MPTP-) lesioned right side substantia nigra of rhesus monkeys with PD, they were found to improve impaired behavior [4]. Lmx1α has been shown to regulate midbrain DA neuronal differentiation, and direct injection of NTN into the midbrain restores degenerating DA neurons [5,6,7,8,9]. The exact mechanism by which BMSCs differentiate into DA neurons remains unclear

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Results

Conclusion