Chonnam Med J.  2011 Apr;47(1):27-30. 10.4068/cmj.2011.47.1.27.

Effects of Sphingosine-1-Phosphate on Neural Differentiation and Neurite Outgrowth in Neuroblastoma Cells

• Lee JH
• Jang S
• Jeong HS
• Park JS

• KMID: 2172241
• DOI: http://doi.org/10.4068/cmj.2011.47.1.27

Abstract

Sphingosine-1-phosphate (S1P) is emerging as a new class of second messenger involved in cellular proliferation, differentiation, and apoptosis and is implicated in diverse physiological functions. Despite many studies on the biological functions of S1P, however, little is known about its role in neuronal differentiation. By use of reverse transcription-polymerase chain reaction and immunostaining, this study aimed to explore whether S1P can differentiate neuroblastoma cells into neural cells. After incubation with 1 uM or 10 uM S1P, the number of neurite-bearing cells increased. Furthermore, the neuroblastoma cells revealed immunoreactivity for neural-specific markers such as GAP43, NFH, and SYP by immunostaining. The expression of NFH, MAP2, SYP, NeuroD1, and SYT mRNA, which is specific for neurons, was increased as shown by RT-PCR studies. The results of this study suggest that that S1P can induce neuronal differentiation and may be a good candidate for the treatment of neurodegenerative diseases.

Keyword

Cell differentiation; Neurons; Sphingosine-1-phosphate

MeSH Terms

Apoptosis
Cell Differentiation
Cell Proliferation
Lysophospholipids
Neurites
Neuroblastoma
Neurodegenerative Diseases
Neurons
RNA, Messenger
Second Messenger Systems
Sphingosine
Lysophospholipids
RNA, Messenger
Sphingosine

Figure

• 

  FIG. 1 Effects of S1P on the number of neurite-bearing cells. After incubation with S1P, neuroblastoma cells revealed morphological changes with neurite outgrowth (top), and the number of neurite-bearing cells was increased dose-dependently (bottom). Magnification is 10×10. Statistical significance is *p<0.05.

• 

  FIG. 2 Immunocytochemical staining for neuron-specific markers in differentiated cells by S1P. Neuroblastoma cells were induced to differentiate into neuronal cells in the presence of S1P. Immunocytochemistry revealed that the expression of neuron-specific markers such as GAP43, NFH, and SYP was increased. Scale bar represents 50 µm (10×10).

• 

  FIG. 3 RT-PCR analysis of neuronal markers in differentiated cells. The expression of NFH, MAP2, SYP, NeuroD1, and SYT mRNA, which is specific for neurons, was increased by treatment with S1P. GAPDH was used as a control.

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