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Cytoskeletal breakdown and apoptosis elicited by NO donors in cerebellar granule cells require NMDA receptor
E Bonfoco1, M Leist, B Zhivotovsky
1Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
Abstract:
We have recently demonstrated that nitric oxide (NO) donors can trigger either apoptosis or necrosis of neurons as a function of the intensity of the exposure. Here, we show that the apoptosis induced by the NO donors S-nitrosocysteine (SNOC) or S-nitroso-N-acetyl-penicillamine (SNAP) in cultured cerebellar granule cells (CGCs) depends on NMDA receptor (NMDA-R) activation leading to intracellular Ca2+ overload. Early dissolution of actin filaments followed by breakdown of microtubules and nuclear lamins preceded the appearance of typical apoptotic features. NO donors induced tyrosine nitration in neurons, in a small population of contaminating astrocytes, and in cultures of cerebellar astroglial cells. However, astrocytes neither displayed cytoskeletal alterations nor underwent apoptosis. Competitive and uncompetitive NMDA receptor antagonists, such as D-aminophosphonovaleric acid and MK-801, did not influence tyrosine nitration but prevented the accumulation of intracellular Ca2+, cytoskeletal breakdown, and apoptosis induced by either SNOC or SNAP in CGCs. Taken together, these data strongly suggest that Ca2+ influx through NMDA-R-gated ion channels is a critical event in CGC apoptosis induced by NO donors.
Insights
Nitric oxide (NO) donors trigger neuronal apoptosis via NMDA receptor activation and calcium overload. This process involves cytoskeletal breakdown, highlighting a critical pathway in NO-induced neurotoxicity.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Nitric oxide (NO) donors can induce neuronal cell death through apoptosis or necrosis, depending on exposure intensity.
- Previous research has established NO's role in neuronal signaling and cell death pathways.
Purpose of the Study:
- To elucidate the specific mechanisms by which NO donors induce apoptosis in cerebellar granule cells (CGCs).
- To investigate the role of NMDA receptors and intracellular calcium in NO-induced neuronal apoptosis.
Main Methods:
- Cultured cerebellar granule cells (CGCs) were treated with NO donors (S-nitrosocysteine and S-nitroso-N-acetyl-penicillamine).
- NMDA receptor antagonists (D-aminophosphonovaleric acid, MK-801) were used to block receptor activity.
- Intracellular calcium levels, cytoskeletal integrity (actin filaments, microtubules, nuclear lamins), and tyrosine nitration were assessed.
Main Results:
- NO donors induced apoptosis in CGCs, characterized by cytoskeletal breakdown and preceded by actin filament dissolution.
- Apoptosis was dependent on NMDA receptor (NMDA-R) activation, leading to intracellular Ca2+ overload.
- NMDA receptor antagonists blocked Ca2+ accumulation, cytoskeletal damage, and apoptosis without affecting tyrosine nitration.
Conclusions:
- Calcium influx through NMDA-R channels is a critical mediator of NO donor-induced apoptosis in CGCs.
- NO-induced neuronal apoptosis involves significant disruption of the neuronal cytoskeleton.
- Astrocytes, despite tyrosine nitration, do not undergo apoptosis, suggesting cell-type-specific responses to NO donors.
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