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. 1994 Feb;111(2):483–488. doi: 10.1111/j.1476-5381.1994.tb14762.x

Fluspirilene block of N-type calcium current in NGF-differentiated PC12 cells.

C J Grantham 1, M J Main 1, M B Cannell 1
PMCID: PMC1909950  PMID: 8004393

Abstract

1. High voltage-activated calcium currents were recorded in nerve growth factor (NGF)-differentiated PC12 cells with the whole-cell patch clamp technique. After exposure to NGF for 3-10 days the PC12 cells developed neurone-like processes and calcium currents which were pharmacologically separable into L- and N-types (defined by sensitivity to nifedipine and omega-conotoxin GVIA respectively). 2. After blocking the L-type calcium channels with nifedipine (10 microM), omega-conotoxin GVIA blocked approximately 85% of the remaining calcium current with an IC50 of 3 nM and a Hill coefficient of 1. The block by conotoxin GVIA was irreversible on the time scale of these experiments. These results suggested that the majority of the nifedipine-insensitive calcium current was N-type. 3. Fluspirilene, a substituted diphenylbutylpiperidine with potent neuroleptic properties, reversibly inhibited the N-type component in a dose-dependent manner with an IC50 of 30 nM. The Hill coefficient of the block was 0.25. The fraction of current blocked was the same at all test potentials examined (-30 to +40 mV). 4. These data indicate that the neuroleptic properties of fluspirilene may be due, at least in part, to an inhibition of neuronal N-type calcium channels. This finding raises the possibility that modulation of N-type calcium channel activity by drugs derived from substituted diphenylbutylpiperidines may provide a novel way of altering neurotransmitter release and hence brain function.

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Selected References

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  1. Doherty P., Mann D. A., Walsh F. S. Comparison of the effects of NGF, activators of protein kinase C, and a calcium ionophore on the expression of Thy-1 and N-CAM in PC12 cell cultures. J Cell Biol. 1988 Jul;107(1):333–340. doi: 10.1083/jcb.107.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Durante W., Sen A. K., Sunahara F. A. Impairment of endothelium-dependent relaxation in aortae from spontaneously diabetic rats. Br J Pharmacol. 1988 Jun;94(2):463–468. doi: 10.1111/j.1476-5381.1988.tb11548.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Enyeart J. J., Biagi B. A., Day R. N., Sheu S. S., Maurer R. A. Blockade of low and high threshold Ca2+ channels by diphenylbutylpiperidine antipsychotics linked to inhibition of prolactin gene expression. J Biol Chem. 1990 Sep 25;265(27):16373–16379. [PubMed] [Google Scholar]
  4. Fox A. P., Nowycky M. C., Tsien R. W. Kinetic and pharmacological properties distinguishing three types of calcium currents in chick sensory neurones. J Physiol. 1987 Dec;394:149–172. doi: 10.1113/jphysiol.1987.sp016864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Furukawa K., Onodera H., Kogure K., Akaike N. Time-dependent expression of Na and Ca channels in PC12 cells by nerve growth factor and cAMP. Neurosci Res. 1993 Feb;16(2):143–147. doi: 10.1016/0168-0102(93)90081-z. [DOI] [PubMed] [Google Scholar]
  6. Galizzi J. P., Fosset M., Romey G., Laduron P., Lazdunski M. Neuroleptics of the diphenylbutylpiperidine series are potent calcium channel inhibitors. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7513–7517. doi: 10.1073/pnas.83.19.7513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gandolfo G., Gottesmann C., Bidard J. N., Lazdunski M. Ca2+ channel blockers prevent seizures induced by a class of K+ channel inhibitors. Eur J Pharmacol. 1989 Jan 24;160(1):173–177. doi: 10.1016/0014-2999(89)90669-9. [DOI] [PubMed] [Google Scholar]
  8. Garber S. S., Hoshi T., Aldrich R. W. Regulation of ionic currents in pheochromocytoma cells by nerve growth factor and dexamethasone. J Neurosci. 1989 Nov;9(11):3976–3987. doi: 10.1523/JNEUROSCI.09-11-03976.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gould R. J., Murphy K. M., Reynolds I. J., Snyder S. H. Antischizophrenic drugs of the diphenylbutylpiperidine type act as calcium channel antagonists. Proc Natl Acad Sci U S A. 1983 Aug;80(16):5122–5125. doi: 10.1073/pnas.80.16.5122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haas S., Beckmann H. Pimozide versus Haloperidol in acute schizophrenia. A double blind controlled study. Pharmacopsychiatria. 1982 Mar;15(2):70–74. doi: 10.1055/s-2007-1019512. [DOI] [PubMed] [Google Scholar]
  11. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  12. Hassel P. Experimental comparison of low doses of 1.5 mg fluspirilene and bromazepam in out-patients with psychovegetative disturbances. Pharmacopsychiatry. 1985 Sep;18(5):297–302. doi: 10.1055/s-2007-1017384. [DOI] [PubMed] [Google Scholar]
  13. Hirning L. D., Fox A. P., McCleskey E. W., Olivera B. M., Thayer S. A., Miller R. J., Tsien R. W. Dominant role of N-type Ca2+ channels in evoked release of norepinephrine from sympathetic neurons. Science. 1988 Jan 1;239(4835):57–61. doi: 10.1126/science.2447647. [DOI] [PubMed] [Google Scholar]
  14. Kenny B. A., Fraser S., Kilpatrick A. T., Spedding M. Selective antagonism of calcium channel activators by fluspirilene. Br J Pharmacol. 1990 Jun;100(2):211–216. doi: 10.1111/j.1476-5381.1990.tb15784.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. King V. F., Garcia M. L., Shevell J. L., Slaughter R. S., Kaczorowski G. J. Substituted diphenylbutylpiperidines bind to a unique high affinity site on the L-type calcium channel. Evidence for a fourth site in the cardiac calcium entry blocker receptor complex. J Biol Chem. 1989 Apr 5;264(10):5633–5641. [PubMed] [Google Scholar]
  16. Lapierre Y. D. A controlled study of penfluridol in the treatment of chronic schizophrenia. Am J Psychiatry. 1978 Aug;135(8):956–959. doi: 10.1176/ajp.135.8.956. [DOI] [PubMed] [Google Scholar]
  17. Llinás R., Sugimori M., Lin J. W., Cherksey B. Blocking and isolation of a calcium channel from neurons in mammals and cephalopods utilizing a toxin fraction (FTX) from funnel-web spider poison. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1689–1693. doi: 10.1073/pnas.86.5.1689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller R. J. Multiple calcium channels and neuronal function. Science. 1987 Jan 2;235(4784):46–52. doi: 10.1126/science.2432656. [DOI] [PubMed] [Google Scholar]
  19. Mintz I. M., Venema V. J., Swiderek K. M., Lee T. D., Bean B. P., Adams M. E. P-type calcium channels blocked by the spider toxin omega-Aga-IVA. Nature. 1992 Feb 27;355(6363):827–829. doi: 10.1038/355827a0. [DOI] [PubMed] [Google Scholar]
  20. Mori Y., Friedrich T., Kim M. S., Mikami A., Nakai J., Ruth P., Bosse E., Hofmann F., Flockerzi V., Furuichi T. Primary structure and functional expression from complementary DNA of a brain calcium channel. Nature. 1991 Apr 4;350(6317):398–402. doi: 10.1038/350398a0. [DOI] [PubMed] [Google Scholar]
  21. Nowycky M. C., Fox A. P., Tsien R. W. Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature. 1985 Aug 1;316(6027):440–443. doi: 10.1038/316440a0. [DOI] [PubMed] [Google Scholar]
  22. Olivera B. M., McIntosh J. M., Cruz L. J., Luque F. A., Gray W. R. Purification and sequence of a presynaptic peptide toxin from Conus geographus venom. Biochemistry. 1984 Oct 23;23(22):5087–5090. doi: 10.1021/bi00317a001. [DOI] [PubMed] [Google Scholar]
  23. Plummer M. R., Logothetis D. E., Hess P. Elementary properties and pharmacological sensitivities of calcium channels in mammalian peripheral neurons. Neuron. 1989 May;2(5):1453–1463. doi: 10.1016/0896-6273(89)90191-8. [DOI] [PubMed] [Google Scholar]
  24. Qar J., Galizzi J. P., Fosset M., Lazdunski M. Receptors for diphenylbutylpiperidine neuroleptics in brain, cardiac, and smooth muscle membranes. Relationship with receptors for 1,4-dihydropyridines and phenylalkylamines and with Ca2+ channel blockade. Eur J Pharmacol. 1987 Sep 11;141(2):261–268. doi: 10.1016/0014-2999(87)90271-8. [DOI] [PubMed] [Google Scholar]
  25. Quirion R., Lafaille F., Nair N. P. Comparative potencies of calcium channel antagonists and antischizophrenic drugs on central and peripheral calcium channel binding sites. J Pharm Pharmacol. 1985 Jun;37(6):437–440. doi: 10.1111/j.2042-7158.1985.tb03033.x. [DOI] [PubMed] [Google Scholar]
  26. Rudy B., Kirschenbaum B., Rukenstein A., Greene L. A. Nerve growth factor increases the number of functional Na channels and induces TTX-resistant Na channels in PC12 pheochromocytoma cells. J Neurosci. 1987 Jun;7(6):1613–1625. doi: 10.1523/JNEUROSCI.07-06-01613.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Seeman P. Brain dopamine receptors. Pharmacol Rev. 1980 Sep;32(3):229–313. [PubMed] [Google Scholar]
  28. Sher E., Clementi F. Omega-conotoxin-sensitive voltage-operated calcium channels in vertebrate cells. Neuroscience. 1991;42(2):301–307. doi: 10.1016/0306-4522(91)90376-y. [DOI] [PubMed] [Google Scholar]
  29. Smith S. J., Augustine G. J. Calcium ions, active zones and synaptic transmitter release. Trends Neurosci. 1988 Oct;11(10):458–464. doi: 10.1016/0166-2236(88)90199-3. [DOI] [PubMed] [Google Scholar]
  30. Streit J., Lux H. D. Calcium current inactivation during nerve-growth-factor-induced differentiation of PC12 cells. Pflugers Arch. 1990 Jun;416(4):368–374. doi: 10.1007/BF00370742. [DOI] [PubMed] [Google Scholar]
  31. Streit J., Lux H. D. Distribution of calcium currents in sprouting PC12 cells. J Neurosci. 1989 Dec;9(12):4190–4199. doi: 10.1523/JNEUROSCI.09-12-04190.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Streit J., Lux H. D. Voltage dependent calcium currents in PC12 growth cones and cells during NGF-induced cell growth. Pflugers Arch. 1987 May;408(6):634–641. doi: 10.1007/BF00581167. [DOI] [PubMed] [Google Scholar]
  33. Usowicz M. M., Porzig H., Becker C., Reuter H. Differential expression by nerve growth factor of two types of Ca2+ channels in rat phaeochromocytoma cell lines. J Physiol. 1990 Jul;426:95–116. doi: 10.1113/jphysiol.1990.sp018128. [DOI] [PMC free article] [PubMed] [Google Scholar]

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