Phenotypic characterization of retinoic acid differentiated SH-SY5Y cells by transcriptional profiling

PLoS One. 2013 May 28;8(5):e63862. doi: 10.1371/journal.pone.0063862. Print 2013.

Abstract

Multiple genetic and environmental factors play a role in the development and progression of Parkinson's disease (PD). The main neuropathological hallmark of PD is the degeneration of dopaminergic (DAergic) neurons in the substantia nigra pars compacta. To study genetic and molecular contributors to the disease process, there is a great need for readily accessible cells with prominent DAergic features that can be used for reproducible in vitro cellular screening. Here, we investigated the molecular phenotype of retinoic acid (RA) differentiated SH-SY5Y cells using genome wide transcriptional profiling combined with gene ontology, transcription factor and molecular pathway analysis. We demonstrated that RA induces a general neuronal differentiation program in SH-SY5Y cells and that these cells develop a predominantly mature DAergic-like neurotransmitter phenotype. This phenotype is characterized by increased dopamine levels together with a substantial suppression of other neurotransmitter phenotypes, such as those for noradrenaline, acetylcholine, glutamate, serotonin and histamine. In addition, we show that RA differentiated SH-SY5Y cells express the dopamine and noradrenalin neurotransmitter transporters that are responsible for uptake of MPP(+), a well known DAergic cell toxicant. MPP(+) treatment alters mitochondrial activity according to its proposed cytotoxic effect in DAergic neurons. Taken together, RA differentiated SH-SY5Y cells have a DAergic-like phenotype, and provide a good cellular screening tool to find novel genes or compounds that affect cytotoxic processes that are associated with PD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Methyl-4-phenylpyridinium / toxicity
  • Biomarkers / metabolism
  • Cell Differentiation / drug effects*
  • Cell Differentiation / genetics*
  • Cell Line
  • Dopamine / metabolism
  • Dopamine Plasma Membrane Transport Proteins / metabolism
  • Dopaminergic Neurons / cytology
  • Dopaminergic Neurons / drug effects
  • Dopaminergic Neurons / metabolism
  • Gene Expression Profiling*
  • Gene Expression Regulation / drug effects
  • Humans
  • Neurotransmitter Agents / metabolism
  • Norepinephrine Plasma Membrane Transport Proteins / metabolism
  • Oligonucleotide Array Sequence Analysis
  • Phenotype
  • Substantia Nigra / cytology
  • Substantia Nigra / drug effects
  • Substantia Nigra / metabolism
  • Tretinoin / pharmacology*

Substances

  • Biomarkers
  • Dopamine Plasma Membrane Transport Proteins
  • Neurotransmitter Agents
  • Norepinephrine Plasma Membrane Transport Proteins
  • Tretinoin
  • 1-Methyl-4-phenylpyridinium
  • Dopamine

Grant support

The authors wish to thank Top Institute Pharma (grant number T5-207), Leiden, The Netherlands (www.tipharma.com) and Stichting Internationaal Parkinson Fonds, The Netherlands (www.parkinsonfonds.nl) for financial support. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.