Characterization of catechol-thioether-induced apoptosis in human SH-SY5Y neuroblastoma cells
Luciana Mosca1, Italo Tempera, Eugenio Lendaro
1Department of Biochemical Sciences, University Sapienza, Roma, Italy. luciana.mosca@uniroma1.it
Abstract:
Recent work has highlighted the involvement of a dopamine derivative, 5-S-cysteinyl-dopamine (CysDA), in neurodegeneration and apoptotic cell death. In this paper we study in further detail the apoptotic process activated by this catechol-thioether derivative of dopamine in SH-SY5Y neuroblastoma cells. CysDA activates a cascade of events by an initial perturbation of Calcium homeostasis in the cell. Cell treatment with the catechol-thioether induces an immediate rise in intracellular Ca(2+) concentration, as demonstrated by a shift in the indo-1 dye emission spectrum, and a sustained high calcium concentration at long times of incubation. Fluorescence microscopy data show that the treatment of cells induces mitochondrial transmembrane potential depolarization, a clear evidence of the onset of apoptotic process. Programmed cell death activation is also demonstrated by cytochrome c release from the mitochondria, by an increased activity of both caspase-8 and -9 and by the poly(ADP-ribose)polymerase (PARP-1) cleavage, yielding the typical 86 kDa fragment due to caspase-3 activity. Overall, our data support the hypothesis that CysDA may induce apoptotic death in neuronal cells, via an initial perturbation of calcium homeostasis in the cytosol.
Insights
5-S-cysteinyl-dopamine (CysDA) triggers neurodegeneration by disrupting calcium homeostasis in neuronal cells. This initiates apoptosis through mitochondrial damage and caspase activation, leading to programmed cell death.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- 5-S-cysteinyl-dopamine (CysDA), a dopamine derivative, has been implicated in neurodegeneration.
- Understanding the precise mechanisms of CysDA-induced cell death is crucial for neuroprotection research.
Purpose of the Study:
- To investigate the detailed apoptotic process activated by CysDA in SH-SY5Y neuroblastoma cells.
- To elucidate the role of calcium homeostasis disruption in CysDA-mediated neurotoxicity.
Main Methods:
- Indo-1 dye fluorescence spectroscopy to monitor intracellular calcium levels.
- Fluorescence microscopy to assess mitochondrial transmembrane potential.
- Measurement of cytochrome c release, caspase activity (caspase-8, -9, -3), and PARP-1 cleavage.
Main Results:
- CysDA treatment caused an immediate and sustained increase in intracellular calcium (Ca2+).
- Mitochondrial depolarization and cytochrome c release were observed, indicating apoptosis onset.
- Increased activity of caspase-8, caspase-9, and caspase-3 (evidenced by PARP-1 cleavage) confirmed programmed cell death.
Conclusions:
- CysDA induces apoptotic death in neuronal cells.
- The primary mechanism involves the initial perturbation of cytosolic calcium homeostasis.
- CysDA represents a potential target for understanding and treating neurodegenerative diseases.
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