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Evaluation of SK-N-SH Cells as a Model for NMDA Receptor Induced Toxicity
Gunnar Goerges1, Paul Disse1,2, Stefan Peischard1
1Institute for Genetics of Heart Diseases (IfGH), Department of Cardiovascular Medicine, University Hospital Münster, Robert-Koch-Str. 45, Münster, Germany.
Background/Aims:
Over the years, the number of patients with neurodegenerative diseases is constantly rising illustrating the need for new neuroprotective drugs. A promising treatment approach is the reduction of excitotoxicity induced by rising (S)-glutamate levels and subsequent NMDA receptor overactivation. To facilitate the search for new NMDA receptor inhibitors neuronal cell models are needed. In this study, we evaluated the suitability of human SK-N-SH cells to serve as a cell model for neurodegeneration induced by NMDA receptor overstimulation.
Methods:
The cytoprotective effect of the unselective NMDA receptor blocker ketamine as well as the GluN2B-selective inhibitor WMS14-10 was evaluated utilizing different cell viability assays, such as endpoint (LDH, CCK-8, DAPI/FACS) and time dependent methods (bioimpedance).
Results:
Non-differentiated as well as differentiated SK-N-SH cells express GluN1 and GluN2B subunits. Furthermore, 50 mM (S)-glutamate led to an instantaneous decrease in cell survival. Only application of unselective channel blocker ketamine could protect differentiated cells against this effect, while the selective inhibitor WMS14-10 did not significantly increase cell survival.
Conclusion:
SK-N-SH cells show an increased sensitivity to (S)-glutamate mediated cytotoxicity with higher differentiation level, that is only partially induced by NMDA receptor overstimulation. Furthermore, we showed that only unselective NMDA receptor inhibition can partially reverse (S)-glutamate-induced toxicity.
Insights
Human SK-N-SH cells can model neurodegeneration. Differentiated cells are sensitive to (S)-glutamate, with only unselective NMDA receptor blockers like ketamine offering protection against excitotoxicity.
Area of Science:
- Neuroscience
- Pharmacology
- Cell Biology
Background:
- Rising incidence of neurodegenerative diseases necessitates novel neuroprotective drugs.
- Excitotoxicity, driven by (S)-glutamate and NMDA receptor overactivation, is a key target for neuroprotection.
- Development of effective NMDA receptor inhibitors requires suitable neuronal cell models.
Purpose of the Study:
- To assess the utility of human SK-N-SH cells as a model for NMDA receptor overstimulation-induced neurodegeneration.
- To investigate the neuroprotective potential of NMDA receptor blockers in this cell model.
Main Methods:
- SK-N-SH cells (differentiated and non-differentiated) were analyzed for GluN1 and GluN2B subunit expression.
- Cell viability was assessed using endpoint assays (LDH, CCK-8, DAPI/FACS) and time-dependent bioimpedance.
- The cytoprotective effects of ketamine (unselective NMDA blocker) and WMS14-10 (GluN2B-selective inhibitor) were evaluated.
Main Results:
- SK-N-SH cells express both GluN1 and GluN2B subunits.
- (S)-glutamate (50 mM) caused rapid cell death.
- Ketamine protected differentiated cells, whereas WMS14-10 did not significantly improve survival.
Conclusions:
- SK-N-SH cells exhibit heightened sensitivity to (S)-glutamate excitotoxicity, particularly when differentiated.
- NMDA receptor overstimulation partially contributes to this observed cytotoxicity.
- Only unselective NMDA receptor inhibition demonstrated partial reversal of (S)-glutamate-induced toxicity in this model.
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