Expression of death-associated protein kinase and recruitment to the tumor necrosis factor signaling pathway following brief seizures.
Death-associated protein (DAP) kinase is calcium-regulated and known to function downstream of death receptors, prompting us to examine its role in the mechanism of seizure-induced neuronal death. Brief seizures were focally evoked in rats, eliciting neuronal death within the CA3 subfield of the hippocampus, and to a lesser extent, cortex. Western blotting confirmed expression of DAP kinase within hippocampus and cortex at the predicted weight of approximately 160 kDa. Immunohistochemistry revealed seizures triggered a significant increase in numbers of DAP kinase-expressing cells within CA3 and cortex, without affecting cell counts within seizure-resistant CA2 or the dentate gyrus. Numbers of DAP kinase-expressing cells were increased in relation to specific patterns of injury-causing seizure activity, electrographically defined. Seizures caused an early increase in DAP kinase binding to actin, and association with calmodulin. Co-immunoprecipitation studies also revealed seizures triggered binding of DAP kinase to the tumor necrosis factor receptor 1 and the Fas-associated death domain protein, commensurate with caspase-8 proteolysis. In contrast, within surviving fields of the hippocampus, DAP kinase interacted with the molecular chaperone 14-3-3. These data suggest DAP kinase is involved in the molecular pathways activated during seizure-induced neuronal death.
Henshall
David C
DC
Robert S. Dow Neurobiology Laboratories, Legacy Clinical Research & Technology Center, 1225 NE 2nd Avenue, Portland, OR 97232, USA. dhenshall@DowNeurobiology.org
Araki
Tomohiro
T
Schindler
Clara K
CK
Shinoda
Sachiko
S
Lan
Jing-Quan
JQ
Simon
Roger P
RP
D048948:14-3-3 Proteins; D048868:Adaptor Proteins, Signal Transducing; D000679:Amygdala / Q000187:drug effects / Q000473:pathology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D051017:Apoptosis Regulatory Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D002452:Cell Count; D002540:Cerebral Cortex / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D064550:Death-Associated Protein Kinases; D004195:Disease Models, Animal; D004569:Electroencephalography; D004789:Enzyme Activation; D053200:Fas-Associated Death Domain Protein; D006624:Hippocampus / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D007608:Kainic Acid; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I; D012640:Seizures / Q000139:chemically induced / Q000145:classification / Q000503:physiopathology*; D015398:Signal Transduction / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D048948:14-3-3 Proteins; D048868:Adaptor Proteins, Signal Transducing; D015703:Antigens, CD; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; C505535:Fadd protein, rat; D053200:Fas-Associated Death Domain Protein; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; D014446:Tyrosine 3-Monooxygenase; D064550:Death-Associated Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D007608:Kainic Acid