Possible involvement of both mitochondria- and endoplasmic reticulum-dependent caspase pathways in rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells

doi:10.1016/S0304-3940(02)00964-3

Neuroscience Letters

Volume 333, Issue 1, 15 November 2002, Pages 25-28

https://doi.org/10.1016/S0304-3940(02)00964-3 Get rights and content

Abstract

Recently, it has been shown that rotenone, a specific inhibitor of mitochondrial complex I, is a useful tool in animal models of Parkinson's disease, but the mechanism of rotenone-induced neuronal death is not fully understood. In human neuroblastoma SH-SY5Y cells, rotenone induced the degradation of procaspases-12, -9 and -3, followed by cleavage of poly (adenosine diphosphate-ribose) polymerase, DNA fragmentation and cell death. Pretreatment with phorbol-12-myristate-13-acetate inhibited the rotenone-induced decrease in procaspases-9 and -3, but not that in procaspase-12. In contrast, benzyloxycarbonyl-Val-Ala-Asp(OCH₃)-CH₂F inhibited the decrease in procaspase-12, but not those in procaspases-9 and -3 in this study. These results suggest that rotenone may induce activation of both mitochondria- and endoplasmic reticulum-dependent caspases in human SH-SY5Y cells.

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Acknowledgements

We thank Mr D. Tsuchiya and Ms K. Nakamura for their technical assistance. The present study was supported in part by the Frontier Research Program (T.T.); Grants-in-Aid (Y.K., T.T., Y.N., S.S.) from the Ministry of Education, Science, Sports and Culture of Japan; and a Grant-in-Aid for the promotion of the advancement of education and research in graduate schools (Special Research) from the Promotion and Mutual Aid Corporation for Private Schools of Japan (T.T.).

References (19)

R. Castellani et al.
Glycoxidation and oxidative stress in Parkinson disease and diffuse Lewy body disease
Brain Res.
(1996)
Y. Imai et al.
Parkin suppresses unfolded protein stress-induced cell death through its E3 ubiquitin-protein ligase activity
J. Biol. Chem.
(2000)
Y. Imai et al.
An unfolded putative transmembrane polypeptide, which can lead to endoplasmic reticulum stress, is a substrate of Parkin
Cell
(2001)
Y. Kitamura et al.
Nitric oxide donor-induced p53-sensitive cell death is enhanced by Bcl-2 reduction in human neuroblastoma cells
Neurochem. Int.
(1998)
P. Li et al.
Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade
Cell
(1997)
R.V. Rao et al.
Coupling endoplasmic reticulum stress to the cell death program: mechanism of caspase activation
J. Biol. Chem.
(2001)
R.V. Rao et al.
Coupling endoplasmic reticulum stress to the cell death program: an Apaf-1-independent intrinsic pathway
J. Biol. Chem.
(2002)
A.H. Schapira et al.
Mitochondrial complex I deficiency in Parkinson's disease
Lancet
(1989)
H. Zou et al.
Apaf-1, a human protein homologous to C, elegans CED-4, participates in cytochrome c-dependent activation of caspase-3
Cell
(1997)

There are more references available in the full text version of this article.

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Earlier we have reported the different susceptibility of the rotenone in different brain areas (Swarnkar et al., 2009) and cerebral damage by diminished mitochondrial enzyme activity and increased ROS levels (Swarnkar et al., 2010). Activation of proapoptotic factor like BAD and caspases have also been reported in rotenone treated neuroblastoma SHSY5Y cells (Kitamura et al., 2002; Watabe and Nakaki, 2004). Rotenone induced neuronal death is reported however, in context to neuronal death its effects on astrocytes are not well studied.
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These pioneering findings suggest that melatonin may be a candidate for the development of novel effective antileishmanial agents either alone or in associations with other drugs.
Isradipine prevents rotenone-induced intracellular calcium rise that accelerates senescence in human neuroblastoma SH-SY5Y cells
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However, the mechanism of RT-induced injury remains elusive. Several studies have reported that RT-induces cell apoptosis in SH-SY5Y cell through mitochondrial dysfunction (Kitamura et al., 2002; Newhouse et al., 2004; Bauereis et al., 2011) and regulation of [Ca2+]i (Ilijic et al., 2011). Ca2+ influx is an energy-required process supplied by ATP that is provided by mitochondria through oxidative phosphorylation or flux of ion gradient (Wilson and Callaway, 2000).
Previous research demonstrated that rotenone (RT) induces neuronal injury partially by increasing intracellular Ca²⁺ concentrations ([Ca²⁺]_i), and inducing oxidative stress, leading to a neurodegenerative disorder. However, the mechanism of RT-induced injury remains elusive. Recent work revealed that Ca²⁺ signaling is important for RT-induced senescence in human neuroblastoma SH-SY5Y cells. In the present study, we found that in SH-SY5Y cells, RT increased [Ca²⁺]_i, senescence associated β-galactosidase activity, aggregation of lipofuscin, production of reactive oxygen species, G1/G0 cell cycle arrest, and activation of p53/p21signaling proteins. In addition, RT decreased the expression of the signaling proteins for cell proliferation and survival, Cyclin-dependent kinase 2 (CDK2), cyclin D1, and Akt. Pretreatment of SH-SY5Y cells with isradipine, an L-type Ca²⁺ channel blocker, or EGTA antagonized these effects of RT. These results suggested that application of isradipine might be a novel approach to prevent RT-induced neurodegenerative disorder such as Parkinson’s disease.
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The α-synuclein protein is a major component of Lewy bodies found in the brains of patients with Parkinson's disease (PD). Recently, α-synuclein 98 (α-syn98), a small isoform of the wild type protein was isolated. The neurotoxicity of this protein was assessed by over-expressing α-syn98 in dopaminergic cells. Enhanced expression of α-syn98 was insufficient to adversely affect the survival of neurons or to promote aggregation of the protein. However, when exposed to rotenone, α-syn98 over-expressing dopaminergic cells demonstrated significantly increased cytotoxicity and aggregate formation. Furthermore, we found enhanced basal ROS production and MDA levels in α-syn98 over-expressing neurons. High basal oxidative stress induced by α-syn98, combined with oxidative stress caused by rotenone treatment, promoted aggregate formation and significantly decreased cell viability. These data indicate that α-syn98 can enhance the susceptibility of dopaminergic neurons to oxidative insults by raising steady-state levels of oxidative stress.
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These results demonstrate that activation of ERK1/2 mediates synphilin-1-induced differentiation. To investigate whether synphilin-1 has a protective role in PD-related insult, we used Rotenone (a classical mitochondrial complex I inhibitor) as a model system since Rotenone causes significant oxidative stress and apoptosis in cell culture (Kitamura et al., 2002; Grivennikova and Vinogradov, 2006). Treatment with Rotenone for 24 h period induced apoptotic cell death in vector control cells at concentrations of 20 to 50 nM (Fig. 5A).
Synphilin-1 is a cytoplasmic protein with unclear function. Synphilin-1 has been identified as an interaction partner of α-synuclein. The interaction between synphilin-1 and α-synuclein has implications in Parkinson's disease. In this study, we stably overexpressed human synphilin-1 in mouse N1E-115 neuroblastoma cells. We found that overexpression of synphilin-1 shortened cell growth doubling time and increased neurite outgrowth. Knockdown of endogenous synphilin-1 caused neuronal toxicity and shortened neurite outgrowth. We further found that synphilin-1 increased activation of the extracellular signal-regulated kinases (ERK1/2) and mediated neurite outgrowth. Rotenone, mitochondrial complex I inhibitor, has been shown previously to induce dopaminergic neurodegeneration and Parkinsonism in rats and Drosophila. We found that Rotenone induced apoptotic cell death in N1E-115 cells via caspase-3 activation and poly (ADP-ribose) polymerase (PARP) cleavage. Overexpression of synphilin-1 significantly reduced Rotenone-induced cell death, caspase-3 activation and PARP cleavage. The results indicate that synphilin-1 displays trophic and protective effects in vitro, suggesting that synphilin-1 may play a protective role in Parkinson's disease (PD) pathogenesis and may lead to a potential therapeutic target for PD intervention.

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Possible involvement of both mitochondria- and endoplasmic reticulum-dependent caspase pathways in rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells

Abstract

Section snippets

Acknowledgements

Brain Res.

J. Biol. Chem.

Cell

Neurochem. Int.

Cell

J. Biol. Chem.

J. Biol. Chem.

Lancet

Cell