Summary
Our previous studies suggested that protein kinase C is involved in the veratridine (an activator of voltage-dependent Na+ channels)-induced phosphorylation and activation of tyrosine hydroxylase as well as the synthesis of catecholamines in adrenal medulla (Uezono et al. 1989). In the present study, we investigated whether treatment of cultured bovine adrenal medullary cells with veratridine causes the accumulation of diacylglycerol, a physiological activator of protein kinase C and the translocation of protein kinase C from cytosol to membrane, a process required for protein kinase C activation. Veratridine (100 μmol/l) increased diacylglycerol level about 2.2 fold in a monophasic manner, with peaking at 5 min and declining toward the basal level within 20 min. Veratridine also increased membrane protein kinase C from 15.6% to 26.9% of total protein kinase C in a timecourse similar to that of diacylglycerol accumulation. Both stimulatory effects of veratridine were inhibited by tetrodotoxin and not observed in Ca2+-free, EGTA-containing medium. Amiloride, an inhibitor of Na+/Ca2+ and Na+/H+ exchange, did not alter veratridine-induced events. These results suggest that veratridine-induced Ca2+ influx contributes to the accumulation of diacylglycerol and the activation of protein kinase C in adrenal medullary cells.
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References
Altin JG, Bradshaw R (1990) Production of 1,2-diacylglycerol in PC12 cells by nerve growth factor and basic fibroblast growth factor. J Neurochem 54:1666–1676
Barid JG, Nahorski SR (1990) Increased intracellular calcium stimulates 3H-inositol polyphosphate accumulation in rat cerebral cortical slices. J Neurochem 54:555–561
Benuck M, Reith MEA, Lajtha A (1989a) Phosphoinositide hydrolysis induced by depolarization and sodium channel activation in mouse cerebrocortical slices. Neuropharmacology 28:847–854
Benuck M, Reith MEA, Lajtha A (1989b) Evidence for the involvement of Na+/Ca2+ exchange in the stimulation of inositol phospholipid hydrolysis by sodium channel activation and depolarization. Eur J Pharmacol 159:187–190
Berridge MJ (1987) Inositol triphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem 56:159–193
Besterman JM, Duronio V, Cuatrecasas P (1986) Rapid formation of diacylglycerol from phosphatidylcholine: a pathway for generation of a second messenger. Proc Natl Acad Sci USA 83:6785–6789
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917
Bocckino SB, Blackmore PF, Wilson PB, Exton JH (1987) Phosphatidate accumulation in hormone-treated hepatocytes via a phospholipase D mechanism. J Biol Chem 262:15309–15315
Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa U, Nishizuka Y (1982) Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem 257:7847–7851
Cooper DR, Hernandez H, Kuo JY, Farese RV (1990) Insulin increases the synthesis of phospholipid and diacylglycerol and protein kinase C activity in rat hepatocytes. Arch Biochem Biophys 276:486–494
Corcoran JJ, Kirshner N (1983) Inhibition of calcium uptake, sodium uptake, and catecholamine secretion by methoxyverapamil (D600) in primary cultures of adrenal medulla cells. J Neurochem 40:1106–1109
Dunn LA, Holz RW (1983) Catecholamine secretion from digitonin-treated adrenal medullary chromaffin cells. J Biol Chem 258:4989–4993
Eberhard DA, Holz RW (1987) Cholinergic stimulation of inositol phosphate formation in bovine adrenal chromaffin cells: distinct nicotinic and muscarinic mechanisms. J Neurochem 49:1634–1643
Eberhard DA, Holz RW (1988) Intracellular Ca2+ activates phospholipase C. Trends Neurosci 11:517–520
El Mestikawy S, Gozlan H, Glowinski J, Hamon M (1985) Characteristics of tyrosine hydroxylase activation by K+-induced depolarization and/or forskolin in rat striatal slices. J Neurochem 45:173–184
Galper JB, Catterall WA (1979) Inhibition of sodium channels by D600. Mol Pharmacol 15:174–178
Griendling KK, Berk BC, Alexander RW (1988) Evidence that Na+/H+ exchange regulates angiotensin II-stimulated diacylglycerol accumulation in vascular smooth muscle cells. J Biol Chem 263:10620–10624
Gusovsky F, Daly JW (1988) Formation of inositol phosphates in synaptoneurosomes of guinea pig brain: stimulatory effects of receptor agonists, sodium channel agents and sodium and calcium ionophores. Neuropharmacology 27:95–105
Gusovsky F, Hollingsworth EB, Daly JW (1986) Regulation of phosphatidylinositol turnover in brain synaptoneurosomes: stimulatory effects of agents that enhance influx of sodium ions. Proc Natl Acad Sci USA 83:3003–3007
Gusovsky F, McNeal ET, Daly JW (1987) Stimulation of phosphoinositide breakdown in brain synaptoneurosomes by agents that activate sodium influx: antagonism by tetrodotoxin, saxitoxin, and cadmium. Mol Pharmacol 32:479–487
Haycock JW Browning MD, Greengard P (1988) Cholinergic regulation of protein phosphorylation in bovine adrenal chromaffin cells. Proc Natl Acad Sci USA 85:1677–1681
Ho AK, Thomas TP, Chik CL, Anderson WB, Klein DC (1988) Protein kinase C: subcellular distribution by increased Ca2+ influx. J Biol Chem 263:9292–9297
Horwitz J (1990) Carbachol and bradykinin increase the production of diacylglycerol from sources other than inositol-containing phospholipids in PC12 cells. J Neurochem 54:983–991
Horwitz J, Perlman RL (1984) Stimulation of DOPA synthesis in the superior cervical ganglion by veratridine. J Neurochem 42:384–389
Huang K-P (1989) The mechanism of protein kinase C activation. Trends Neurosci 12:425–432
Irving HR, Exton JH (1987) Phosphatidylcholine breakdown in rat liver plasma membranes. J Biol Chem 262:3440–3443
Kaczorowski GJ, Barros F, Dethmers JK, Trumble MJ (1985) Inhibition of Na+/Ca2+ exchange in pituitary plasma membrane vesicles by analogues of amiloride. Biochemistry 24:1394–1403
Kao L-S, Cheung NS (1990) Mechanism of calcium transport across the plasma membrane of bovine chromaffin cells. J Neurochem 54:1972–1979
Kendall DA, Nahorski SR (1984) Inositol phospholipid hydrolysis in rat cerebral cortical slices: II. calcium requirement. J Neurochem 42:1388–1394
Knight DE, Baker PF (1983) Stimulus-secretion coupling in isolated adrenal medullary cells. QJ Exp Physiol 68:123–143
McDonough PM, Goldstein D, Brown JH (1988) Elevation of cytoplasmic calcium concentration stimulates hydrolysis of phosphatidylinositol bisphosphate in chick heart cells: effect of sodium channel activators. Mol Pharmacol 33:310–315
Nishizuka Y (1986) Studies and perspectives of protein kinase C. Science 233:305–312
Pelech SL, Vance DE (1989) Signal transduction via phosphatidylcholine cycles. Trends Neurosci 14:28–30
Preiss J, Loomis CR, Bishop WR, Stein R, Niedel JE, Bell RM (1986) Quantitative measurement of sn-1,2-diacylglycercols present in platelets, hepatocytes, and ras- and sis-transformed normal rat kidney cells. J Biol Chem 261:8597–8600
Saltiel AR, Sherline P, Fox JA (1987) Insulin-stimulated diacylglycerol production results from the hydrolysis of a novel phosphatidylinositol glycan. J Biol Chem 262:1116–1121
Sasakawa N, Nakaki T, Yamamoto S, Kato R (1989) Calcium uptake-dependent and -independent mechanisms of inositol trisphosphate formation in adrenal chromaffin cells: comparative studies with high K+, carbamylcholine and angiotensin II. Cell Signalling 1:75–84
Terbush DR, Holz RW (1986) Effects of phorbol esters, diglyceride, and cholinergic agonists on the subcellular distribution of protein kinase C in intact or digitonin-permeabilized adrenal medullary cells. J Biol Chem 261:17099–17106
Terbush DR, Bittner MA, Holz RW (1988) Ca2+ influx causes rapid translocation of protein kinase C to membranes. J Biol Chem 263:18 873–18 879
Uezono Y, Yanagihara N, Wada A, Koda Y, Yokota K, Kobayashi H, Izumi F (1989) Veratridine-induced phosphorylation and activation of tyrosine hydroxylase, and synthesis of catecholamines in cultured bovine adrenal medullary cells. Naunyn-Schmiedeberg's Arch Pharmacol 339:653–659
Wada A, Izumi F, Yanagihara N, Kobayashi H (1985 a) Modulation by ouabain and diphenylhydantoin of veratridine-induced 22Na influx and its relation to 45Ca influx and the secretion of catecholamines in cultured bovine adrenal medullary cells. Naunyn-Schmiedeberg's Arch Pharmacol 328:273–278
Wada A, Takara H, Izumi F, Kobayashi H, Yanagihara N (1985 b) Influx of 22Na through acetylcholine receptor-associated Na channels: relationship between 22Na influx, 45Ca influx and secretion of catecholamines in cultured bovine adrenal medulla cells. Neuroscience 15:283–292
Wolf M, Cuatrecasas P, Sahyoun N (1985) Interaction of protein kinase C with membranes is regulated by Ca2+, phorbol esters, and ATP. J Biol Chem 260:15718–15722
Wu WC-S, Walaas SI, Nairn AC, Greengard P (1982) Calcium/phospholipid regulates phosphorylation of a Mr “87K” substrate protein in brain synaptosomes. Proc Natl Acad Sci USA 79:5249–5253
Yanagihara N, Yokota K, Kobayashi H, Wada A, Uezono Y, Izumi F (1990) Sodium/proton exchange in cultured bovine adrenal medullarry cells. J Neurochem 54:1626–1631
Yatani A, Brown AM (1985) The calcium channel blocker nitrendipine blocks sodium channels in neonatal rat cardiac myocytes. Circ Res 56:868–875
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Uezono, Y., Wada, A., Yanagihara, N. et al. Veratridine causes the Ca2+-dependent increase in diacylglycerol formation and translocation of protein kinase C to membranes in cultured bovine adrenal medullary cells. Naunyn-Schmiedeberg's Arch Pharmacol 346, 76–81 (1992). https://doi.org/10.1007/BF00167574
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DOI: https://doi.org/10.1007/BF00167574