Peptide inhibitors of caspase-3-like proteases attenuate MPP+ -induced toxicity of cultured fetal rat mesencephalic dopamine neurons

doi:10.1016/S0306-4522(98)00154-7

Neuroscience

Volume 86, Issue 3, 8 June 1998, Pages 701-707

https://doi.org/10.1016/S0306-4522(98)00154-7 Get rights and content

Abstract

Multiple aspartate-specific cysteine proteases have been identified and specific members of this family have been implicated in the apoptotic death of many mammalian cell types. Caspase-3-like proteases seem to play a pivotal role in neuronal apoptosis since mice with germline inactivation of the caspase-3 gene manifest profound alterations in neurogenesis. Moreover, inhibitors of caspase-3-related proteases have been shown to inhibit neuronal apoptosis. Here we extend recent work from our laboratory on the mechanisms mediating the neurotoxic actions of 1-phenyl-4-methylpyridinium using ventral mesencephalon cultures containing dopamine neurons. We demonstrate that low concentrations of 1-phenyl-4-methylpyridinium induce apoptosis in dopamine neurons by morphological and biochemical criteria. Moreover, pretreatment of ventral mesencephalon cultures with the tetrapeptide inhibitors of the caspase-3-like proteases; zVAD-FMK or Ac-DEVD-CHO specifically inhibit death of dopamine neurons neurons induced by low concentrations of 1-phenyl-4-methylpyridinium, whereas the caspase-1-like inhibitor Ac-YVAD-CHO was without effect. Our data indicate that exposure of cultured ventral mesencephalondopamine neurons to low concentrations of 1-phenyl-4-methylpyridinium results in apoptotic death and that caspase-3-like proteases may mediate the neurotoxic apoptotic actions of 1-phenyl-4-methylpyridinium.

References (37)

E.S Alnemri et al.
Human ICE/CED-3 protease nomenclature
Cell
(1996)
M Ankarcrona et al.
Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function
Neuron
(1995)
B Dipasquale et al.
Apoptosis and DNA degradation induced by 1-methyl-pyridinium in neurons
Biochem. biophys. Res. Commun.
(1991)
Y Itano et al.
1-methyl-4-phenyl-pyridinium ion (MPP⁺) causes fragmentation and increases the Bcl-2 expression in human neuroblastoma, SH-SY5Y cells, through different mechanisms
Brain Res.
(1995)
P Li et al.
Mice deficient in IL-1ß-converting enzyme are defective in production of mature IL-1ß and resistant to endotoxic shock
Cell
(1995)
N Margolin et al.
Substrate and inhibitor specivity of interleukin-1ß-converting enzyme and related caspases
J. biol. Chem.
(1997)
H Mochizuki et al.
Apoptosis is induced by 1-methyl-pyridinium ion (MPP⁺) in ventral mesencephalic-striatal coculture in rat
Neurosci. Lett.
(1994)
T Mutoh et al.
1-methyl-4-pyridinium kills differented PC12 cells with a concomitant change in protein phosphorylation
Brain Res.
(1994)
D.W Nicholson et al.
Caspases: killer proteases
Trends biochem. Sci.
(1997)
J.C Reed
Cytochrome c: can't live with it
Cell
(1997)

R.V Talanian et al.

Substrate specifities of caspase family proteases

J. biol. Chem.

(1997)

N.A Tatton et al.

In situ detection of apoptotic nuclei in the substantia nigra compacta of 1-methyl-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal deoxynucleotidyl transferase labelling and acridine orange staining

Neuroscience

(1997)

G.M Yan et al.

Depolarization or glutamate receptor activation blocks apoptotic cell death of cultured cerebellar granule neurons

Brain Res.

(1994)

J Yuan et al.

The C. elegans cell death gene ced-3 encodes a protein similar to mammalian interleukin-1ß-converting enzyme

Cell

(1993)

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)

P Anglade et al.

Apoptosis and autophagy in nigral neurons of patients with Parkinson's disease

Histol. Histopathol.

(1997)

R.C Armstrong et al.

Activation of the CED3/ICE-related protease CPP32 in cerebellar granule neurons undergoing apoptosis but not necrosis

J. Neurosci.

(1997)

K.E Bowenkamp et al.

Glial cell line-derived neurotrophic factor supports survival of injured midbrain dopaminergic neurons

J. comp. Neurol.

(1995)

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¹: These authors contributed equally to this work.

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Letter to NeurosciencePeptide inhibitors of caspase-3-like proteases attenuate MPP+ -induced toxicity of cultured fetal rat mesencephalic dopamine neurons

Abstract

Cell

Neuron

Biochem. biophys. Res. Commun.

Brain Res.

Cell

J. biol. Chem.

Neurosci. Lett.

Brain Res.

Trends biochem. Sci.

Cell

J. biol. Chem.

Neuroscience

Brain Res.

Cell

Cell

Apoptosis and autophagy in nigral neurons of patients with Parkinson's disease

Histol. Histopathol.

Activation of the CED3/ICE-related protease CPP32 in cerebellar granule neurons undergoing apoptosis but not necrosis

J. Neurosci.

Glial cell line-derived neurotrophic factor supports survival of injured midbrain dopaminergic neurons

J. comp. Neurol.

Letter to Neuroscience
Peptide inhibitors of caspase-3-like proteases attenuate MPP⁺ -induced toxicity of cultured fetal rat mesencephalic dopamine neurons