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TNF-alpha stimulates caspase-3 activation and apoptotic cell death in primary septo-hippocampal cultures.
Zhao, X; Bausano, B; Pike, B R; Newcomb-Fernandez, J K; Wang, K K; Shohami, E; Ringger, N C; DeFord, S M; Anderson, D K; Hayes, R L.
Afiliação
  • Zhao X; The Vivian L. Smith Center for Neurologic Research, Department of Neurosurgery, The University of Texas Health Science Center, Houston, Texas, USA.
J Neurosci Res ; 64(2): 121-31, 2001 Apr 15.
Article em En | MEDLINE | ID: mdl-11288141
Primary septo-hippocampal cell cultures were incubated in varying concentrations of tumor necrosis factor (TNF-alpha; 0.3-500 ng/ml) to examine proteolysis of the cytoskeletal protein alpha-spectrin (240 kDa) to a signature 145 kDa fragment by calpain and to the apoptotic-linked 120-kDa fragment by caspase-3. The effects of TNF-alpha incubation on morphology and cell viability were assayed by fluorescein diacetate-propidium iodide (FDA-PI) staining, assays of lactate dehydrogenase (LDH) release, nuclear chromatin alterations (Hoechst 33258), and internucleosomal DNA fragmentation. Incubation with varying concentrations of TNF-alpha produced rapid increases in LDH release and nuclear PI uptake that were sustained over 48 hr. Incubation with 30 ng/ml TNF-alpha yielded maximal, 3-fold, increase in LDH release and was associated with caspase-specific 120-kDa fragment but not calpain-specific 145-kDa fragment as early as 3.5 hr after injury. Incubation with the pan-caspase inhibitor, carbobenzosy- Asp-CH(2)-OC (O)-2-6-dichlorobenzene (Z-D-DCB, 50-140 microM) significantly reduced LDH release produced by TNF-alpha. Apoptotic-associated oligonucleosomal-sized DNA fragmentation on agarose gels was detected from 6 to 72 hr after exposure to TNF-alpha. Histochemical changes included chromatin condensation, nuclear fragmentation, and formation of apoptotic bodies. Results of this study suggest TNF-alpha may induce caspase-3 activation but not calpain activation in septo-hippocampal cultures and that this activation of caspase-3 at least partially contributes to TNF-alpha-induced apoptosis.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Septo Pelúcido / Fator de Necrose Tumoral alfa / Apoptose / Caspases / Hipocampo / Proteínas do Tecido Nervoso / Neurônios Limite: Animals Idioma: En Revista: J Neurosci Res Ano de publicação: 2001 Tipo de documento: Article País de afiliação: Estados Unidos País de publicação: Estados Unidos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Septo Pelúcido / Fator de Necrose Tumoral alfa / Apoptose / Caspases / Hipocampo / Proteínas do Tecido Nervoso / Neurônios Limite: Animals Idioma: En Revista: J Neurosci Res Ano de publicação: 2001 Tipo de documento: Article País de afiliação: Estados Unidos País de publicação: Estados Unidos