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Robust Dopaminergic Differentiation and Enhanced LPS-Induced Neuroinflammatory Response in Serum-Deprived Human SH-SY5Y Cells: Implication for Parkinson’s Disease

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Abstract

Parkinson’s disease (PD) is a chronic neurodegenerative condition characterized by motor symptoms such as bradykinesia, resting tremor, and rigidity. PD diagnosis is based on medical history, review of signs, symptoms, neurological and physical examinations. Unfortunately, by the time the disease is diagnosed, dopamine (DA) neuronal loss is often extended, thereby resulting in ineffective therapies. Recent evidence suggests that neuroinflammation may be pivotal during PD onset and progression. However, suitable cellular models and biomarkers to detect early signs of neuroinflammation are still missing. In this study, we developed a well-differentiated DAergic neuronal cell line where we triggered a neuroinflammatory response to assess the temporal expression of the tissue- and urokinase plasminogen activators (tPA and uPA) and their endogenous inhibitor (PAI-1) along with that of pro-inflammatory mediators and the neuronal marker nNOS. Human neuroblastoma cells SH-SY5Y were differentiated into DAergic neuronal-like cells using a combination of 12-O-tetradecanoylphorbol-13-acetate (TPA) and serum depletion. Terminally-differentiated neurons were then exposed to lipopolysaccharide (LPS) for short (up to 24 h) or long term (up to 10 days) to mimic acute or chronic inflammation. Results demonstrated that uPA protein expression was stably upregulated during chronic inflammation, whereas the expression of nNOS protein better reflected the cellular response to acute inflammation. Additional studies revealed that the temporal induction of uPA was associated with increased AKT phosphorylation, but did not seem to involve cAMP-responsive element-binding protein (CREB) activation, nor the mitogen-activated protein kinase (MAPK) pathway. In conclusion, our in vitro data suggests that nNOS and uPA may serve as viable candidate biomarkers of acute and chronic neuroinflammation.

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Acknowledgments

We would like to acknowledge A/Prof. Louise Cole and the UTS Microscopy Imaging Facility (MIF) for the support provided for the acquisition and processing of the immunofluorescent images shown in this study.

Funding

These experiments were supported by the University of Technology Sydney Start-up Research Fund 2018 to Dr. Castorina.

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Authors and Affiliations

  1. Laboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, PO Box 123, Broadway, NSW, 2007, Australia

    Aram Niaz, Jocelyn Karunia, Mawj Mandwie, Ghaith Al-Badri & Alessandro Castorina

  2. Laboratory of Neural Structure and Function (LNSF), School of Medical Sciences, (Anatomy & Histology), Faculty of Medicine and Health, University of Sydney, Sydney, NSW, 2006, Australia

    Kevin A. Keay & Alessandro Castorina

  3. Anatomy, Histology and Movement Sciences Section, Department of Biomedical and Biotechnological Sciences, School of Medicine, University of Catania, via S. Sofia, 87, Catania, 95123, Italy

    Giuseppe Musumeci

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  1. Aram Niaz
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  2. Jocelyn Karunia
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Correspondence to Alessandro Castorina.

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Niaz, A., Karunia, J., Mandwie, M. et al. Robust Dopaminergic Differentiation and Enhanced LPS-Induced Neuroinflammatory Response in Serum-Deprived Human SH-SY5Y Cells: Implication for Parkinson’s Disease. J Mol Neurosci 71, 565–582 (2021). https://doi.org/10.1007/s12031-020-01678-6

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