Effects of receptor antagonists, Ca2+ channel blocker, Ca2+depletion, and protein kinase inhibitors on CREB phosphorylation in cultured hippocampal neurons.

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Effects of receptor antagonists, Ca2+ channel blocker, Ca2+depletion, and protein kinase inhibitors on CREB phosphorylation in cultured hippocampal neurons after exposure to glutamate. Effects of receptor antagonists, Ca2+ channel blocker, Ca2+depletion, and protein kinase inhibitors on CREB phosphorylation in cultured hippocampal neurons after exposure to glutamate. As indicated, reagents used in this study were MK-801 (10 μm), nifedipine (10 μm), Ca-free medium with CA2+-chelator EGTA (1 mm), Rp-8-cAMPs (100 μm), KT5720 (2 μm), chelerythrine (10 μm), Rp-8-cGMPs (100 μm), KN93 (30 μm), PD98059 (50 μm), genistein (200 μm), wortmannin (400 nm), and staurosporine (100 nm). Cultured neurons were treated for 30 min before the addition of and during the 10 min exposure to 100 μmglutamate. Enhanced CREB phosphorylation induced by exposure to glutamate was prevented in the Ca2+-free condition and in the presence of MK-801, KN93, and staurosporine as indicated in the graphical quantification. Data are mean ± SD (n = 6 for each treatment); *p < 0.05 versus control. Takuma Mabuchi et al. J. Neurosci. 2001;21:9204-9213 ©2001 by Society for Neuroscience