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Aim:To investigate the changes in synchronized spontaneous Ca~(2+)oscillationsinduced by mitogen-activated protein kinase kinase(MEK)inhibitor PD98059 atdifferent concentrations in cultured hippocampal network.Methods:Hippocam-pal neurons in culture for 1-2 weeks were used for this study.Spontaneoussynaptic activities of these hippocampal neurons were examined by Ca~(2+)imagingusing calcium-sensitive dye.MEK inhibitor PD98059(10,30,and 60μmol/L)andSB202474(10 and 60μmol/L),a negative control for mitogen-activated proteinkinase(MAPK)cascade study,were applied to the cells under the microscopewhile imaging was taking place.Results:PD98059 at a lower concentration of 10μmol/L had little effect on the Ca~(2+)oscillation.At the higher concentration of 30μmol/L,5 min after application of PD98059,the spike frequency was decreased to25.38%±7.40%(mean±SEM,n=16,P<0.01 vs medium control)of that of the controlperiod.At an even higher concentration of 60μmol/L,5 min after application ofPD98059,the spike frequency was decreased to 14.53%±5.34%(mean±SEM,n=16,P<0.01 vs medium control)of that of the control period.The spike amplitudeunderwent a corresponding decrease.However,the negative control SB202474 atconcentrations of 10 and 60 μmol/L had little inhibition effect on the Ca~(2+)oscillation.Conclusion:These results indicate that PD98059 inhibits synchronized sponta-neous Ca~(2+)oscillation through inhibition of MEK,which hints that the MAPKcascade is required to maintain synchronized spontaneous Ca~(2+)oscillation.
Aim: To investigate the changes in synchronized spontaneous Ca ~ (2+) oscillations induced by mitogen-activated protein kinase kinase (MEK) inhibitor PD98059 at different concentrations in cultured hippocampal network. Methods: Hippocam-pal neurons in culture for 1-2 weeks were used for this study. Spontaneoussynaptic activities of these hippocampal neurons were examined by Ca 2+ imaging using calcium-sensitive dye. MEK inhibitor PD98059 (10,30, and 60 μmol / L) and SB202474 (10 and 60 μmol / L) for mitogen-activated proteinkinase (MAPK) cascade study, were applied to the cells under the microscope while imaging was taking place. Results: PD98059 at a lower concentration of 10 μmol / L had little effect on the Ca ~ (2+) oscillation. At the the higher concentration of 30 μmol / L for 5 min after application of PD98059, the spike frequency was decreased to 25.38% ± 7.40% (mean ± SEM, n = 16, P <0.01 vs. medium control) of that control period. Atan even higher concentration of 60 μmol / L, 5 min after application of PD980 59, the spike frequency was decreased to 14.53% ± 5.34% (mean ± SEM, n = 16, P <0.01 vs medium control) of that of the control period. The negative control SB202474 atconcentrations of 10 and 60 μmol / L had little inhibition effect on the Ca ~ (2 +) oscillation. Conclusion: These results indicate that PD98059 inhibits synchronized sponta-neous Ca 2+ through through inhibition of MEK, which hints that the MAPK cascade is required to maintain the synchronized spontaneous Ca ~ (2 +) oscillation.