The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
권호 표지

Enhancement of ATP-induced Currents by Phospholipase D1 Overexpressed in PC12 Cells

  • Park, Jin-Bong (Departments of Physiology, College of Medicine, Chungnam National University) ;
  • Kim, Young-Rae (Departments of Biochemistry, College of Medicine, Chungnam National University) ;
  • Jeon, Byeong-Hwa (Departments of Physiology, College of Medicine, Chungnam National University) ;
  • Park, Seung-Kiel (Departments of Biochemistry, College of Medicine, Chungnam National University) ;
  • Oh, Sae-Ock (Departments of Biochemistry, College of Medicine, Chungnam National University) ;
  • Kim, Young-Geun (Departments of Physiology, College of Medicine, Chungnam National University) ;
  • Lee, Sang-Do (Departments of Physiology, College of Medicine, Chungnam National University) ;
  • Kim, Kwang-Jin (Departments of Physiology, College of Medicine, Chungnam National University)
  • Published : 2003.08.21
Download PDF
⟨ Previous Next ⟩

Abstract

Using phospholipase D1 (PLD1)-overexpressing PC12 (PLD1-PC12) cells, the regulatory roles of PLD1 on ATP-induced currents were investigated. In control and PLD1-PC12 cells, ATP increased PLD activity in an external $Ca^{2+}$ dependent manner. PLD activity stimulated by ATP was substantially larger in PLD1-PC12 cells than in control cells. In whole-cell voltage-clamp mode, ATP induced transient inward and outward currents. The outward currents inhibited by TEA or charybdotoxin were significantly larger in PLD1-PC12 cells than in control cells. The inward currents known as $Ca^{2+}$ permeable nonselective cation currents were also larger in PLD1-PC12 cells than in control cells. However, the difference between the two groups of cells disappeared in $Ca^{2+}$-free external solution, where ATP did not activate PLD. Finally, ATP-induced $^{45}Ca$ uptakes were also larger in PLD1-PC12 cells than in control cells. These results suggest that PLD enhances ATP-induced $Ca^{2+}$ influx via $Ca^{2+}$ permeable nonselective cation channels and increases subsequent $Ca^{2+}$-activated $K^+$ currents in PC12 cells.

Keywords

References

  1. Exton JH. Phospholipase D: enzymology, mechanisms of regulation, and function. Physiol Rev 77:303－320, 1997 https://doi.org/10.1152/physrev.1997.77.2.303
  2. Fasolato C, Pizzo P, Pozzan P. Receptor-mediated calcium influx in PC12 cells. ATP and bradykinin activate two independent pathways. J Biol Chem 265:20351－20355, 1990
  3. Fujii F, Kimura J, Tase C. Ca2$^+$-activated K$^+$ current induced by external ATP in PC12 cells. Clin Exp Pharmacol Physiol 26: 39－ 47, 1999 https://doi.org/10.1046/j.1440-1681.1999.02980.x
  4. Gibbs TC, Meier KE. Expression and regulation of phospholipase D isoforms in mammalian cell lines. J Cell Physiol 182:77－87, 2000 https://doi.org/10.1002/(SICI)1097-4652(200001)182:1<77::AID-JCP9>3.0.CO;2-B
  5. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pfl$\"{u}$gers Arch 391:85 －100, 1981 https://doi.org/10.1007/BF00656997
  6. Hayakawa K, Nakashima S, Ito Y, Mizuta K, Miyata H, Nozawa Y. Increased expression of phospholipase D1 mRNA during cAMP- or NGF-induced differentiation in PC12 cells. Neurosci Lett 265:127－130, 1999 https://doi.org/10.1016/S0304-3940(99)00228-1
  7. Hoshi T, Aldrich RW. Voltage-dependent K$^+$ currents and underlying single K$^+$ channels in pheochromocytoma cells. J Gen Physiol 91:73－106, 1988 https://doi.org/10.1085/jgp.91.1.73
  8. Humeau Y, Vitale N, Chasserot-Golaz S, Dupont JL, Du G, Frohman MA, Bader MF, Poulain B. A role for phospholipase D1 in neurotransmitter release. Proc Natl Acad Sci USA 98 (26):15300－15305, 2001 https://doi.org/10.1073/pnas.261358698
  9. Ito Y, Nakashima S, Nozawa Y. Hydrogen peroxide-induced phospholipase D activation in rat pheochromocytoma PC12 cells: possible involvement of Ca2$^+$-dependent protein tyrosine kinase. J Neurochem 69:729－736, 1997 https://doi.org/10.1046/j.1471-4159.1997.69020729.x
  10. Jordan M, Schallhorn A, Wurm FM. Transfecting mammalian cells: optimization of critical parameters affecting calcium-phosphate precipitate formation. Nucleic Acids Res 24:596－601, 1996 https://doi.org/10.1093/nar/24.4.596
  11. Khiroug L, Giniatullin R, Talantova M, Nistri A. Role of intracellular calcium in fast and slow desensitization of P2-receptors in PC12 cells. Br J Pharmacol 120 (8):1552－1560, 1997 https://doi.org/10.1038/sj.bjp.0701060
  12. Kim WK, Rabin RA. Characterization of the purinergic P2 receptors in PC12 cells. Evidence for a novel subtype. J Biol Chem 269:6471－6477, 1994
  13. Knabb MT, Rubio R, Berne RM. Calcium-dependent atrial slow action potentials generated with phosphatidic acid or phospholipase D. Pfl$\"{u}$gers Arch 401:435－437, 1984 https://doi.org/10.1007/BF00584352
  14. Latorre R, Oberhauser A, Labarca P, Alvarez O. Varieties of calcium-activated potassium channels. Ann Rev Physiol 51: 385 －399, 1989 https://doi.org/10.1146/annurev.ph.51.030189.002125
  15. Min DS, Ahn BH, Rhie DJ, Yoon SH, Hahn SJ, Kim MS, Jo YH. Expression and regulation of phospholipase D during neuronal differentiation of PC12 cells. Neuropharmacology 41:384－391, 2001 https://doi.org/10.1016/S0028-3908(01)00070-3
  16. Nakazawa K, Fujimori K, Takanaka A, Inoue K. An ATP-activated conductance in pheochromocytoma cells and its suppression by extracellular calcium. J Physiol 428: 257－272, 1990 https://doi.org/10.1113/jphysiol.1990.sp018211
  17. Nakazawa K, Hess P. Block by calcium of ATP-activated channels in pheochromocytoma cells. J Gen Physiol 101:377－392, 1993
  18. Nakazawa K, Inoue K. Roles of Ca2$^+$ influx through ATP-activated channels in catecholamine release from pheochromocytoma PC12 cells. J Neurophysiol 68:2026－2032, 1992 https://doi.org/10.1152/jn.1992.68.6.2026
  19. Oh SO, Hong JH, Kim YR, Yoo HS, Lee SH, Lim K, Hwang BD, Exton JH, Park SK. Regulation of phospholipase D2 by H2O2 in PC12 cells. J Neurochem 75:2445－2454, 2000
  20. Park JB, Kim HJ, Ryu PD, Moczydlowski E. Effect of Phosphatidylserine on Unitary Conductance and Ba2$^+$ Block of the BK Ca2$^+$-activated K$^+$ Channel: Re-examination of the Surface Charge Hypothesis. J Gen Physiol 121 (5):375－398, 2003 https://doi.org/10.1085/jgp.200208746
  21. Sah P. Ca2$^+$-activated K$^+$ currents in neurones: types, physiological roles and modulation. Trends Neurosci 19:150－154, 1996 https://doi.org/10.1016/S0166-2236(96)80026-9
  22. Schmid-Antomarchi H, Hugues M, Lazdunski M. Properties of the apamin-sensitive Ca2$^+$-activated K$^+$ channel in PC12 pheochro mocytoma cells which hyper-produce the apamin receptor. J Biol Chem 261:8633－8637, 1986
  23. Sela D, Ram E, Atlas D. ATP receptor. A putative receptor-operated channel in PC-12 cells. J Biol Chem 266:17990－17994, 1991
  24. Vitale N, Caumont AS, Chasserot-Golaz S, Du G, Wu S, Sciorra VA, Morris AJ, Frohman MA, Bader MF. Phospholipase D1: a key factor for the exocytotic machinery in neuroendocrine cells. EMBO J 20:2424－2434, 2001 https://doi.org/10.1093/emboj/20.10.2424
  25. Vitale N, Chasserot-Golaz S, Bader MF. Regulated secretion in chromaffin cells: an essential role for ARF6-regulated phospholipase D in the late stages of exocytosis. Ann N Y Acad Sci 971: 193－200, 2002 https://doi.org/10.1111/j.1749-6632.2002.tb04463.x
  26. Walter M, Tepel M, Nofer JR, Neusser M, Assmann G, Zidek W. Involvement of phospholipase D in store-operated calcium influx in vascular smooth muscle cells. FEBS Lett 479:51－56, 2000 https://doi.org/10.1016/S0014-5793(00)01880-9