Key Proteins of Activating Cell Death Can Be Predicted through a Kainic Acid-Induced Excitotoxic Stress

Epilepsy is a major neurological disorder characterized by spontaneous seizures accompanied by neurophysiological changes. Repeated seizures can damage the brain as neuronal death occurs. A better understanding of the mechanisms of brain cell death could facilitate the discovery of novel treatments for neurological disorders such as epilepsy. In this study, a model of kainic acid- (KA-) induced neuronal death was established to investigate the early protein markers associated with apoptotic cell death due to excitotoxic damage in the rat cortex. The results indicated that KA induces both apoptotic and necrotic cell death in the cortex. Incubation with high concentrations (5 and 500 μM, >75%) and low concentrations (0.5 pM: 95% and 50 nM: 8%) of KA for 180 min led to necrotic and apoptotic cell death, respectively. Moreover, proteomic analysis using two-dimensional gel electrophoresis and mass spectrometry demonstrated that antiapoptotic proteins, including heat shock protein 70, 3-mercaptopyruvate sulfurtransferase, tubulin-B-5, and pyruvate dehydrogenase E1 component subunit beta, were significantly higher in apoptosis than in necrosis induced by KA. Our findings provide direct evidence that several proteins are associated with apoptotic and necrotic cell death in excitotoxicity model. The results indicate that these proteins can be apoptotic biomarkers from the early stages of cell death.

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Concurrent Assessment of Calpain and Caspase-3 Activation after Oxygen–Glucose Deprivation in Primary Septo-Hippocampal Cultures

Journal of Cerebral Blood Flow & Metabolism ◽

10.1097/00004647-200111000-00004 ◽

2001 ◽

Vol 21 (11) ◽

pp. 1281-1294 ◽

Cited By ~ 47

Author(s):

Jennifer K. Newcomb-Fernandez ◽

Xiurong Zhao ◽

Brian R. Pike ◽

Kevin K. W. Wang ◽

Andreas Kampfl ◽

...

Keyword(s):

Cell Death ◽

Caspase 3 ◽

Apoptotic Cell ◽

Glucose Deprivation ◽

Apoptotic Cell Death ◽

Oxygen Glucose Deprivation ◽

Necrotic Cell Death ◽

Necrotic Cell ◽

Lactate Dehydrogenase Release ◽

Hippocampal Cultures

The contributions of calpain and caspase-3 to apoptosis and necrosis after central nervous system (CNS) trauma are relatively unexplored. No study has examined concurrent activation of calpain and caspase-3 in necrotic or apoptotic cell death after any CNS insult. Experiments used a model of oxygen–glucose deprivation (OGD) in primary septo-hippocampal cultures and assessed cell viability, occurrence of apoptotic and necrotic cell death phenotypes, and protease activation. Immunoblots using an antibody detecting calpain and caspase-3 proteolysis of α-spectrin showed greater accumulation of calpain-mediated breakdown products (BDPs) compared with caspase-3–mediated BDPs. Administration of calpain and caspase-3 inhibitors confirmed that activation of these proteases contributed to cell death, as inferred by lactate dehydrogenase release. Oxygen–glucose deprivation resulted in expression of apoptotic and necrotic cell death phenotypes, especially in neurons. Immunocytochemical studies of calpain and caspase-3 activation in apoptotic cells indicated that these proteases are almost always concurrently activated during apoptosis. These data demonstrate that calpain and caspase-3 activation is associated with expression of apoptotic cell death phenotypes after OGD, and that calpain activation, in combination with caspase-3 activation, could contribute to the expression of apoptotic cell death by assisting in the degradation of important cellular proteins.

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Caspase-3-associated apoptotic cell death in excitotoxic necrosis of the entorhinal cortex following intraperitoneal injection of kainic acid in the rat

Neuroscience Letters ◽

10.1016/s0304-3940(01)02518-6 ◽

2002 ◽

Vol 321 (3) ◽

pp. 182-186 ◽

Cited By ~ 44

Author(s):

B. Puig ◽

I. Ferrer

Keyword(s):

Cell Death ◽

Kainic Acid ◽

Entorhinal Cortex ◽

Intraperitoneal Injection ◽

Caspase 3 ◽

Apoptotic Cell ◽

Apoptotic Cell Death

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RING-finger protein 166 plays a novel pro-apoptotic role in neurotoxin-induced neurodegeneration via ubiquitination of XIAP

Cell Death and Disease ◽

10.1038/s41419-020-03145-x ◽

2020 ◽

Vol 11 (10) ◽

Author(s):

Chang-Ki Oh ◽

Young Ki Choi ◽

Ih-Yeon Hwang ◽

Yeon Uk Ko ◽

In Kwon Chung ◽

...

Keyword(s):

Cell Death ◽

Neuronal Death ◽

Apoptotic Cell ◽

Neuronal Cells ◽

Ring Finger ◽

Experimental Models ◽

Apoptotic Cell Death ◽

Ring Finger Protein ◽

Caspase Activation ◽

Finger Protein

Abstract The dopaminergic neurotoxin, 6-hydroxydopamine (6-OHDA), has been widely utilized to establish experimental models of Parkinson disease and to reveal the critical molecules and pathway underlying neuronal death. The profile of gene expression changes following 6-OHDA treatment of MN9D dopaminergic neuronal cells was investigated using a TwinChip Mouse-7.4K microarray. Functional clustering of altered sets of genes identified RING-finger protein 166 (RNF166). RNF166 is composed of an N-terminal RING domain and C-terminal ubiquitin interaction motif. RNF166 localized in the cytosol and nucleus. At the tissue level, RNF166 was widely expressed in the central nervous system and peripheral organs. In the cerebral cortex, its expression decreased over time. In certain conditions, overexpression of RNF166 accelerates the naturally occurring neuronal death and 6-OHDA–induced MN9D cell death as determined by TUNEL and annexin-V staining, and caspase activation. Consequently, 6-OHDA–induced apoptotic cell death was attenuated in RNF166-knockdown cells. In an attempt to elucidate the mechanism underlying this pro-apoptotic activity, binding protein profiles were assessed using the yeast two-hybrid system. Among several potential binding candidates, RNF166 was shown to interact with the cytoplasmic X-linked inhibitor of apoptosis (XIAP), inducing ubiquitin-dependent degradation of XIAP and eventually accelerating caspase activation following 6-OHDA treatment. RNF166’s interaction with and resulting inhibition of the XIAP anti-caspase activity was further enhanced by XIAP-associated factor-1 (XAF-1). Consequently, depletion of RNF166 suppressed 6-OHDA–induced caspase activation and apoptotic cell death, which was reversed by XIAP knockdown. In summary, our data suggest that RNF166, a novel E3 ligase, plays a pro-apoptotic role via caspase activation in neuronal cells.

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Differential bradykinin B1 and B2 receptor regulation in cell death induced by hepatic ischaemia/reperfusion injury

Clinical Science ◽

10.1042/cs20130313 ◽

2014 ◽

Vol 127 (6) ◽

pp. 405-413 ◽

Cited By ~ 2

Author(s):

Mayra A. Paio ◽

Maria Kouyoumdjian ◽

Durval R. Borges ◽

Marcia R. Nagaoka

Keyword(s):

Cell Death ◽

Reperfusion Injury ◽

Apoptotic Cell ◽

Receptor Regulation ◽

Necrotic Cell Death ◽

Necrotic Cell ◽

Ischaemia Reperfusion Injury ◽

B1 Receptor ◽

Ischaemia Reperfusion ◽

Kinin B1 Receptor

In the present study, we have demonstrated that the kinin B1 receptor may participate in apoptotic cell death signalling, whereas the B2 receptor may be involved in necrotic cell death during IRI.

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