TY - JOUR
T1 - AMPA-induced dark cell degeneration of cerebellar Purkinje neurons involves activation of caspases and apparent mitochondrial dysfunction
AU - Strahlendorf, Jean
AU - Box, Cathy
AU - Attridge, Jennifer
AU - Diertien, Janet
AU - Finckbone, VelvetLee
AU - Henne, William M.
AU - Medina, Margarita S.
AU - Miles, Randy
AU - Oomman, Sowmini
AU - Schneider, Marcia
AU - Singh, Hema
AU - Veliyaparabil, Madhu
AU - Strahlendorf, Howard
N1 - Funding Information:
This research was supported in part by a DOD grant and a Howard Hughes Medical Institute grant through the Undergraduate Biological Sciences Education Program to Texas Tech University.
PY - 2003/12/24
Y1 - 2003/12/24
N2 - Cerebellar Purkinje neurons (PNs) are selectively vulnerable to AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepriopionic acid)-induced delayed neurotoxicity known as dark cell degeneration (DCD) that is expressed as cytoplasmic and nuclear condensation, neuron shrinkage, and failure of physiology. The present study was initiated to determine whether AMPA-receptor-induced DCD in PNs is associated with Bax translocation to the mitochondria, cytochrome C release from the mitochondria, changes in mitochondrial potential, and activation of representative initiator and executor caspases that include caspase-9, caspase-3, and caspase-7. AMPA consistently and rapidly hyperpolarized mitochondria as reflected by an increase in MitoTracker Red CMS Ros fluorescence. Increases in Bax immunoreactivity were quantitatively and temporally variable and Bax failed to localize to mitochondria. Additionally, we observed a marked increase in immunoreactivity of cytochrome C although its release from mitochondria was not apparent. Mitochondrial membrane hyperpolarization and increases in cytochrome C immunoreactivity preceded caspase activation. Immunohistochemical analyses revealed the active form of caspases-3 and -9 were markedly and significantly increased in PNs following 30 μM AMPA, and caspase-9 activation preceded caspase-3. Increases in active caspase-7 immunoreactivity were less frequently encountered in PNs. Thus DCD shares some characteristics of apoptotic programmed cell death, but lacks typical mitochondrial pathophysiology associated with classic apoptosis. These findings suggest that AMPA-induced DCD is a form of active PCD that lies on a spectrum between classical apoptosis and passive necrosis.
AB - Cerebellar Purkinje neurons (PNs) are selectively vulnerable to AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepriopionic acid)-induced delayed neurotoxicity known as dark cell degeneration (DCD) that is expressed as cytoplasmic and nuclear condensation, neuron shrinkage, and failure of physiology. The present study was initiated to determine whether AMPA-receptor-induced DCD in PNs is associated with Bax translocation to the mitochondria, cytochrome C release from the mitochondria, changes in mitochondrial potential, and activation of representative initiator and executor caspases that include caspase-9, caspase-3, and caspase-7. AMPA consistently and rapidly hyperpolarized mitochondria as reflected by an increase in MitoTracker Red CMS Ros fluorescence. Increases in Bax immunoreactivity were quantitatively and temporally variable and Bax failed to localize to mitochondria. Additionally, we observed a marked increase in immunoreactivity of cytochrome C although its release from mitochondria was not apparent. Mitochondrial membrane hyperpolarization and increases in cytochrome C immunoreactivity preceded caspase activation. Immunohistochemical analyses revealed the active form of caspases-3 and -9 were markedly and significantly increased in PNs following 30 μM AMPA, and caspase-9 activation preceded caspase-3. Increases in active caspase-7 immunoreactivity were less frequently encountered in PNs. Thus DCD shares some characteristics of apoptotic programmed cell death, but lacks typical mitochondrial pathophysiology associated with classic apoptosis. These findings suggest that AMPA-induced DCD is a form of active PCD that lies on a spectrum between classical apoptosis and passive necrosis.
KW - AMPA
KW - Caspases
KW - Cerebellum
KW - Excitotoxicity
KW - Mitochondria
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U2 - 10.1016/j.brainres.2003.09.048
DO - 10.1016/j.brainres.2003.09.048
M3 - Article
C2 - 14642640
AN - SCOPUS:10744230059
SN - 0006-8993
VL - 994
SP - 146
EP - 159
JO - Brain Research
JF - Brain Research
IS - 2
ER -