title: Concurrent assessment of calpain and caspase-3 activation after oxygen-glucose deprivation in primary septo-hippocampal cultures.
abstract: 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 alpha-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.
authors.affiliation: Department of Neurosurgery, The Vivian L. Smith Center for Neurologic Research, University of Texas Health Science Center, Houston, Texas, USA.
authors.forename: J K
authors.initials: JK
authors.lastname: Newcomb-Fernandez
authors.forename: X
authors.initials: X
authors.lastname: Zhao
authors.forename: B R
authors.initials: BR
authors.lastname: Pike
authors.forename: K K
authors.initials: KK
authors.lastname: Wang
authors.forename: A
authors.initials: A
authors.lastname: Kampfl
authors.forename: R
authors.initials: R
authors.lastname: Beer
authors.forename: S M
authors.initials: SM
authors.lastname: DeFord
authors.forename: R L
authors.initials: RL
authors.lastname: Hayes
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D015153:Blotting, Western; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005674:Fusobacterium Infections; D005947:Glucose / Q000494:pharmacology*; D006624:Hippocampus / Q000166:cytology; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D010100:Oxygen / Q000494:pharmacology*; D010641:Phenotype; D051381:Rats; D015427:Reperfusion Injury / Q000201:enzymology / Q000473:pathology; D012397:Rosette Formation; D020665:Septum of Brain / Q000166:cytology; D013049:Spectrin / Q000378:metabolism; D020521:Stroke / Q000201:enzymology / Q000473:pathology
chemical_list: D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D013049:Spectrin; D002154:Calpain; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D005947:Glucose; D010100:Oxygen