Abstract
Aim:
To investigate the changes in synchronized spontaneous Ca2+ oscillations induced by mitogen-activated protein kinase kinase (MEK) inhibitor PD98059 at different concentrations in cultured hippocampal network.
Methods:
Hippocampal neurons in culture for 1-2 weeks were used for this study. Spontaneous synaptic activities of these hippocampal neurons were examined by Ca2+ imaging using calcium-sensitive dye. MEK inhibitor PD98059 (10, 30, and 60 μmol/L) and SB202474 (10 and 60 μmol/L), a negative control for mitogen-activated protein kinase (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 Ca2+ oscillation. At the higher concentration of 30 μmol/L, 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 of the control period. At an even higher concentration of 60 μmol/L, 5 min after application of PD98059, 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 amplitude underwent a corresponding decrease. However, the negative control SB202474 at concentrations of 10 and 60 μmol/L had little inhibition effect on the Ca2+ oscillation.
Conclusion:
These results indicate that PD98059 inhibits synchronized spontaneous Ca2+ oscillation through inhibition of MEK, which hints that the MAPK cascade is required to maintain synchronized spontaneous Ca2+ oscillation.
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Seger R, Krebs EG . The MAPK signaling cascade. FASEB J 1995; 9: 726–35.
Lazou A, Sugden PH, Clerk A . Activation of mitogen-activated protein kinases (p38-MAPKs, SAPKs/JNKs and ERKs) by the G-protein-coupled receptor agonist phenylephrine in the perfused rat heart. Biochem J 1998; 332: 459–65.
Haq SE, Clerk A, Sugden PH . Activation of mitogen-activated protein kinases (p38-MAPKs, SAPKs/JNKs and ERKs) by adenosine in the perfused rat heart. FEBS Lett 1998; 434: 305–8.
Bogoyevitch MA, Gillespie-Brown J, Ketterman AJ, Fuller SJ, Ben-Levy R, Ashworth A, et al. Stimulation of the stress-activated mitogen-activated protein kinase subfamilies in perfused heart. p38/RK mitogen-activated protein kinases and c-Jun N-terminal kinases are activated by ischemia/reperfusion. Circ Res 1996; 79: 162–73.
Stofega MR, Yu CL, Wu J, Jove R . Activation of extracellular signal-regulated kinase (ERK) by mitogenic stimuli is repressed in v-Src-transformed cells. Cell Growth Differ 1997; 8: 113–9.
Marais R, Marshall CJ . Control of the ERK MAP kinase cascade by Ras and Raf. Cancer Surv 1996; 27: 101–25.
Harrisingh MC, Perez-Nadales E, Parkinson DB, Malcolm DS, Mudge AW, Lloyd AC . The Ras/Raf/ERK signalling pathway drives Schwann cell dedifferentiation. EMBO J 2004; 23: 3061–71.
Boulton TG, Cobb MH . Identification of multiple extracellular signal-regulated kinases (ERKs) with antipeptide antibodies. Cell Regul 1991; 2: 357–71.
Thomas KL, Hunt SP . The regional distribution of extracellularly regulated kinase-1 and -2 messenger RNA in the adult rat central nervous system. Neuroscience 1993; 56: 741–57.
Fiore RS, Bayer VE, Pelech SL, Posada J, Cooper JA, Baraban JM . p42 mitogen-activated protein kinase in brain: prominent localization in neuronal cell bodies and dendrites. Neuroscience 1993; 55: 463–72.
Alessi DR, Cuenda A, Cohen P, Dudley DT, Saltiel AR . PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J Biol Chem 1995; 270: 27489–94.
English JD, Sweatt JD . A requirement for the mitogen-activated protein kinase cascade in hippocampal long term potentiation. J Biol Chem 1997; 272: 19103–6.
Coogan AN, O'Leary DM, O'Connor JJ . P42/44 MAP kinase inhibitor PD98059 attenuates multiple forms of synaptic plasticity in rat dentate gyrus in vitro. J Neurophysiol 1999; 81: 103–10.
Wang X, Gruenstein EI . Mechanism of synchronized Ca2+ oscillations in cortical neurons. Brain Res 1997; 767: 239–49.
Tanaka T, Saito H, Matsuki N . Intracellular calcium oscillation in cultured rat hippocampal neurons: a model for glutamatergic neurotransmission. Jpn J Pharmacol 1996; 70: 89–93.
Liu XH, Lu GW, Cui ZJ . Calcium oscillations in freshly isolated neonatal rat cortical neurons. Acta Pharmacol Sin 2002; 23: 577–81.
Koizumi S, Inoue K . Inhibition by ATP of calcium oscillations in rat cultured hippocampal neurones. Br J Pharmacol 1997; 122: 51–8.
Lisman JE . Bursts as a unit of neural information: making unreliable synapses reliable. Trends Neurosci 1997; 20: 38–43.
Bacci A, Verderio C, Pravettoni E, Matteoli M . Synaptic and intrinsic mechanisms shape synchronous oscillations in hippocampal neurons in culture. Eur J Neurosci 1999; 11: 389–97.
Traub RD, Wong RK . Cellular mechanism of neuronal synchronization in epilepsy. Science 1982; 216: 745–7.
Miles R, Wong RK . Single neurones can initiate synchronized population discharge in the hippocampus. Nature 1983; 306: 371–3.
Ansari HR, Husain S, Abdel-Latif AA . Activation of p42/p44 mitogen-activated protein kinase and contraction by prostaglandin F2alpha, ionomycin, and thapsigargin in cat iris sphincter smooth muscle: inhibition by PD98059, KN-93, and isoproterenol. J Pharmacol Exp Ther 2001; 299: 178–86.
Kupzig S, Walker SA, Cullen PJ . The frequencies of calcium oscillations are optimized for efficient calcium-mediated activation of Ras and the ERK/MAPK cascade. Proc Natl Acad Sci USA 2005; 102: 7577–82.
Doherty P, Williams G, Williams EJ . CAMs and axonal growth: a critical evaluation of the role of calcium and the MAPK cascade. Mol Cell Neurosci 2000; 16: 283–95.
Banker GA, Cowan WM . Rat hippocampal neurons in dispersed cell culture. Brain Res 1977; 126: 397–42.
Gee KR, Brown KA, Chen WN, Bishop-Stewart J, Gray D, Johnson I . Chemical and physiological characterization of Fluo-4 Ca(2+)-indicator dyes. Cell Calcium 2000; 27: 97–106.
Leinekugel X, Medina I, Khalilov I, Ben-Ari Y, Khazipov R . Ca2+ oscillations mediated by the synergistic excitatory actions of GABA(A) and NMDA receptors in the neonatal hippocampus. Neuron 1997; 18: 243–55.
Dudley DT, Pang L, Decker SJ, Bridges AJ, Saltiel AR . A synthetic inhibitor of the mitogen-activated protein kinase cascade. Proc Natl Acad Sci USA 1995; 92: 7686–9.
Brewer GJ, Torricelli JR, Evege EK, Price PJ . Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J Neurosci Res 1993; 35: 567–76.
Pereira DB, Carvalho AP, Duarte CB . Non-specific effects of the MEK inhibitors PD098,059 and U0126 on glutamate release from hippocampal synaptosomes. Neuropharmacology 2002; 42: 9–19.
Gould MC, Stephano JL . MAP kinase, meiosis, and sperm centrosome suppression in Urechis caupo. Dev Biol 1999; 216: 175–9.
Gould MC, Stephano JL . Inactivation of Ca(2+) action potential channels by the MEK inhibitor PD98059. Exp Cell Res 2000; 260: 175–9.
Gu X, Spitzer NC . Distinct aspects of neuronal differentiation encoded by frequency of spontaneous Ca2+ transients. Nature 1995; 375: 784–7.
English JD, Sweatt JD . Activation of p42 mitogen-activated protein kinase in hippocampal long term potentiation. J Biol Chem 1996; 271: 24329–32.
Pisani A, Bonsi P, Martella G, De Persis C, Costa C, Pisani F, et al. Intracellular calcium increase in epileptiform activity: modulation by levetiracetam and lamotrigine. Epilepsia 2004; 45: 719–28.
Shang ZC, Sun BZ, Chen ZN, Wang J, Feng Q, Wang W, et al. Effect of PD98059 on Ras-MAPK signal transduction pathway of chronic myelogenous leukemia. Xi Bao Yu Fen Zi Mian Yi Xue ZaZhi 2003; 19: 54–5.
Yaari Y, Konnerth A, Heinemann U . Spontaneous epileptiform activity of CA1 hippocampal neurons in low extracellular calcium solutions. Exp Brain Res 1983; 51: 153–6.
Murray B, Alessandrini A, Cole AJ, Yee AG, Furshpan EJ . Inhibition of the p44/42 MAP kinase pathway protects hippocampal neurons in a cell-culture model of seizure activity. Proc Natl Acad Sci USA 1998; 95: 11975–80.
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Rui, Yf., Sun, Zh., Gu, Jp. et al. MEK inhibitor PD98059 acutely inhibits synchronized spontaneous Ca2+ oscillations in cultured hippocampal networks. Acta Pharmacol Sin 27, 869–876 (2006). https://doi.org/10.1111/j.1745-7254.2006.00387.x
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DOI: https://doi.org/10.1111/j.1745-7254.2006.00387.x
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