Calcium-dependent expression of TNF-alpha in neural cells is mediated by the calcineurin/NFAT pathway.
We report induction of TNF-alpha via the calcium/calcineurin/NFAT pathway in PC12 neural cells. In PC12, expression of TNF-alpha mRNA, protein and TNF-alpha gene promoter activity was induced by co-stimulation with phorbol ester and either calcium ionophore A23187 or the L-type Voltage Gated Calcium Channel agonist Bay K 8644. Pre-treatment with calcineurin inhibitors CsA or FK506 inhibited the dominant calcium-dependent component of this induction, limiting it to the level achieved with phorbol ester alone. Promoter activation by Bay was abolished by nifedipine, a specific inhibitor of L-type Voltage Gated Calcium Channels. Exogenous NFAT protein transactivated the TNF-alpha promoter, and the peptide VIVIT-a specific inhibitor of calcineurin/NFAT binding-blocked calcium-inducible transactivation of the TNF-alpha promoter. Given proposed functions of TNF-alpha in spatial learning, memory and the pathogenesis of neurodegenerative diseases, the data presented suggest an important role for calcineurin/NFAT signaling in these key neurological processes.
Canellada
Andrea
A
Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas (CBM-CSIC), Universidad Autónoma de Madrid (UAM), Facultad de Ciencias, Madrid 28049, Spain.
Cano
Eva
E
Sánchez-Ruiloba
Lucía
L
Zafra
Francisco
F
Redondo
Juan Miguel
JM
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D019703:Calcineurin / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002120:Calcium Channel Agonists / Q000494:pharmacology; D020013:Calcium Signaling / Q000502:physiology; D003513:Cycloheximide / Q000494:pharmacology; D016572:Cyclosporine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005786:Gene Expression Regulation; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology; D050778:NFATC Transcription Factors / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D016716:PC12 Cells; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D016559:Tacrolimus / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*
D002120:Calcium Channel Agonists; D004791:Enzyme Inhibitors; D007166:Immunosuppressive Agents; D007476:Ionophores; D050778:NFATC Transcription Factors; D009842:Oligopeptides; D011500:Protein Synthesis Inhibitors; D014409:Tumor Necrosis Factor-alpha; C121611:VIVIT peptide; D000001:Calcimycin; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D016572:Cyclosporine; D003513:Cycloheximide; D019703:Calcineurin; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium; D016559:Tacrolimus