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. 1987 Nov;84(21):7739–7743. doi: 10.1073/pnas.84.21.7739

Activators of protein kinase C enhance acetylcholine receptor desensitization in sympathetic ganglion neurons.

J E Downing 1, L W Role 1
PMCID: PMC299376  PMID: 3478722

Abstract

Recent studies suggest that phosphorylation may regulate the rate of desensitization of nicotinic acetylcholine (AcCho) receptors (AcChoR) in vertebrate muscle and Torpedo. It is not known if phosphorylation is involved in regulation of the neuronal AcChoR, however. In this study we examine the possibility that protein kinase C might regulate nicotinic AcChoR function in neurons. Several activators of protein kinase C (1-oleoyl-2-acetylglycerol, phorbol 12,13-diacetate, and phorbol 12,13-dibutyrate) were tested for their ability to modulate AcChoR function in embryonic chicken sympathetic ganglion neurons. Neurons were voltage-clamped at the resting potential, and the response to AcCho was tested before and after treatment with activators of protein kinase C. We find that all of these agents enhance the rate of decay of AcCho-induced current without affecting peak current amplitude or cellular input resistance. The drugs were ineffective if applied concurrently with AcCho: significant effects could be detected after 60 sec of pretreatment. A phorbol that does not increase protein kinase C activity (4 beta-phorbol) was ineffective in enhancing the decay of AcCho-induced current. Thus, the effects of these agents on AcChoR function are likely to be mediated by their interaction with C kinase, rather than by direct interaction with the AcChoR channel. Our data suggest that kinase C may regulate agonist-induced desensitization of the neuronal AcChoR channel.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams P. R., Feltz A. End-plate channel opening and the kinetics of quinacrine (mepacrine) block. J Physiol. 1980 Sep;306:283–306. doi: 10.1113/jphysiol.1980.sp013397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albuquerque E. X., Deshpande S. S., Aracava Y., Alkondon M., Daly J. W. A possible involvement of cyclic AMP in the expression of desensitization of the nicotinic acetylcholine receptor. A study with forskolin and its analogs. FEBS Lett. 1986 Apr 7;199(1):113–120. doi: 10.1016/0014-5793(86)81235-2. [DOI] [PubMed] [Google Scholar]
  3. Boulter J., Evans K., Goldman D., Martin G., Treco D., Heinemann S., Patrick J. Isolation of a cDNA clone coding for a possible neural nicotinic acetylcholine receptor alpha-subunit. 1986 Jan 30-Feb 5Nature. 319(6052):368–374. doi: 10.1038/319368a0. [DOI] [PubMed] [Google Scholar]
  4. Boyd N. D., Leeman S. E. Multiple actions of substance P that regulate the functional properties of acetylcholine receptors of clonal rat PC12 cells. J Physiol. 1987 Aug;389:69–97. doi: 10.1113/jphysiol.1987.sp016647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Browning M. D., Huganir R., Greengard P. Protein phosphorylation and neuronal function. J Neurochem. 1985 Jul;45(1):11–23. doi: 10.1111/j.1471-4159.1985.tb05468.x. [DOI] [PubMed] [Google Scholar]
  6. Castagna M., Takai Y., Kaibuchi K., Sano K., Kikkawa U., Nishizuka Y. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847–7851. [PubMed] [Google Scholar]
  7. Choi D. W., Fischbach G. D. GABA conductance of chick spinal cord and dorsal root ganglion neurons in cell culture. J Neurophysiol. 1981 Apr;45(4):605–620. doi: 10.1152/jn.1981.45.4.605. [DOI] [PubMed] [Google Scholar]
  8. Clapham D. E., Neher E. Substance P reduces acetylcholine-induced currents in isolated bovine chromaffin cells. J Physiol. 1984 Feb;347:255–277. doi: 10.1113/jphysiol.1984.sp015065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DeRiemer S. A., Strong J. A., Albert K. A., Greengard P., Kaczmarek L. K. Enhancement of calcium current in Aplysia neurones by phorbol ester and protein kinase C. Nature. 1985 Jan 24;313(6000):313–316. doi: 10.1038/313313a0. [DOI] [PubMed] [Google Scholar]
  10. Eusebi F., Molinaro M., Zani B. M. Agents that activate protein kinase C reduce acetylcholine sensitivity in cultured myotubes. J Cell Biol. 1985 Apr;100(4):1339–1342. doi: 10.1083/jcb.100.4.1339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldman D., Deneris E., Luyten W., Kochhar A., Patrick J., Heinemann S. Members of a nicotinic acetylcholine receptor gene family are expressed in different regions of the mammalian central nervous system. Cell. 1987 Mar 27;48(6):965–973. doi: 10.1016/0092-8674(87)90705-7. [DOI] [PubMed] [Google Scholar]
  12. Gordon A. S., Davis C. G., Milfay D., Diamond I. Phosphorylation of acetylcholine receptor by endogenous membrane protein kinase in receptor-enriched membranes of Torpedo californica. Nature. 1977 Jun 9;267(5611):539–540. doi: 10.1038/267539a0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hammond C., Paupardin-Tritsch D., Nairn A. C., Greengard P., Gerschenfeld H. M. Cholecystokinin induces a decrease in Ca2+ current in snail neurons that appears to be mediated by protein kinase C. 1987 Feb 26-Mar 4Nature. 325(6107):809–811. doi: 10.1038/325809a0. [DOI] [PubMed] [Google Scholar]
  15. Higashida H., Brown D. A. Two polyphosphatidylinositide metabolites control two K+ currents in a neuronal cell. 1986 Sep 25-Oct 1Nature. 323(6086):333–335. doi: 10.1038/323333a0. [DOI] [PubMed] [Google Scholar]
  16. Horwitz J., Anderson C. H., Perlman R. L. Comparison of the effects of muscarine and vasopressin on inositol phospholipid metabolism in the superior cervical ganglion of the rat. J Pharmacol Exp Ther. 1986 Apr;237(1):312–317. [PubMed] [Google Scholar]
  17. Huganir R. L., Delcour A. H., Greengard P., Hess G. P. Phosphorylation of the nicotinic acetylcholine receptor regulates its rate of desensitization. Nature. 1986 Jun 19;321(6072):774–776. doi: 10.1038/321774a0. [DOI] [PubMed] [Google Scholar]
  18. Huganir R. L., Greengard P. cAMP-dependent protein kinase phosphorylates the nicotinic acetylcholine receptor. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1130–1134. doi: 10.1073/pnas.80.4.1130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Huganir R. L., Miles K., Greengard P. Phosphorylation of the nicotinic acetylcholine receptor by an endogenous tyrosine-specific protein kinase. Proc Natl Acad Sci U S A. 1984 Nov;81(22):6968–6972. doi: 10.1073/pnas.81.22.6968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kaibuchi K., Takai Y., Sawamura M., Hoshijima M., Fujikura T., Nishizuka Y. Synergistic functions of protein phosphorylation and calcium mobilization in platelet activation. J Biol Chem. 1983 Jun 10;258(11):6701–6704. [PubMed] [Google Scholar]
  21. Kaibuchi K., Takai Y., Sawamura M., Hoshijima M., Fujikura T., Nishizuka Y. Synergistic functions of protein phosphorylation and calcium mobilization in platelet activation. J Biol Chem. 1983 Jun 10;258(11):6701–6704. [PubMed] [Google Scholar]
  22. Kandel E. R., Schwartz J. H. Molecular biology of learning: modulation of transmitter release. Science. 1982 Oct 29;218(4571):433–443. doi: 10.1126/science.6289442. [DOI] [PubMed] [Google Scholar]
  23. Kikkawa U., Nishizuka Y. The role of protein kinase C in transmembrane signalling. Annu Rev Cell Biol. 1986;2:149–178. doi: 10.1146/annurev.cb.02.110186.001053. [DOI] [PubMed] [Google Scholar]
  24. Levitan I. B. Phosphorylation of ion channels. J Membr Biol. 1985;87(3):177–190. doi: 10.1007/BF01871217. [DOI] [PubMed] [Google Scholar]
  25. Malenka R. C., Madison D. V., Andrade R., Nicoll R. A. Phorbol esters mimic some cholinergic actions in hippocampal pyramidal neurons. J Neurosci. 1986 Feb;6(2):475–480. doi: 10.1523/JNEUROSCI.06-02-00475.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Margiotta J. F., Berg D. K. Enkephalin and substance P modulate synaptic properties of chick ciliary ganglion neurons in cell culture. Neuroscience. 1986 May;18(1):175–182. doi: 10.1016/0306-4522(86)90186-7. [DOI] [PubMed] [Google Scholar]
  27. McHugh E. M., McGee R., Jr Direct anesthetic-like effects of forskolin on the nicotinic acetylcholine receptors of PC12 cells. J Biol Chem. 1986 Mar 5;261(7):3103–3106. [PubMed] [Google Scholar]
  28. Middleton P., Jaramillo F., Schuetze S. M. Forskolin increases the rate of acetylcholine receptor desensitization at rat soleus endplates. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4967–4971. doi: 10.1073/pnas.83.13.4967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mizobe F., Kozousek V., Dean D. M., Livett B. G. Pharmacological characterization of adrenal paraneurons: substance P and somatostatin as inhibitory modulators of the nicotinic response. Brain Res. 1979 Dec 14;178(2-3):555–566. doi: 10.1016/0006-8993(79)90714-5. [DOI] [PubMed] [Google Scholar]
  30. Nestler E. J., Walaas S. I., Greengard P. Neuronal phosphoproteins: physiological and clinical implications. Science. 1984 Sep 21;225(4668):1357–1364. doi: 10.1126/science.6474180. [DOI] [PubMed] [Google Scholar]
  31. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  32. Rane S. G., Dunlap K. Kinase C activator 1,2-oleoylacetylglycerol attenuates voltage-dependent calcium current in sensory neurons. Proc Natl Acad Sci U S A. 1986 Jan;83(1):184–188. doi: 10.1073/pnas.83.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Role L. W., Leeman S. E., Perlman R. L. Somatostatin and substance P inhibit catecholamine secretion from isolated cells of guinea-pig adrenal medulla. Neuroscience. 1981;6(9):1813–1821. doi: 10.1016/0306-4522(81)90215-3. [DOI] [PubMed] [Google Scholar]
  34. Role L. W. Substance P modulation of acetylcholine-induced currents in embryonic chicken sympathetic and ciliary ganglion neurons. Proc Natl Acad Sci U S A. 1984 May;81(9):2924–2928. doi: 10.1073/pnas.81.9.2924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schuetze S. M., Role L. W. Developmental regulation of nicotinic acetylcholine receptors. Annu Rev Neurosci. 1987;10:403–457. doi: 10.1146/annurev.ne.10.030187.002155. [DOI] [PubMed] [Google Scholar]
  36. Shuster M. J., Camardo J. S., Siegelbaum S. A., Kandel E. R. Cyclic AMP-dependent protein kinase closes the serotonin-sensitive K+ channels of Aplysia sensory neurones in cell-free membrane patches. 1985 Jan 31-Feb 6Nature. 313(6001):392–395. doi: 10.1038/313392a0. [DOI] [PubMed] [Google Scholar]
  37. Siegelbaum S. A., Camardo J. S., Kandel E. R. Serotonin and cyclic AMP close single K+ channels in Aplysia sensory neurones. Nature. 1982 Sep 30;299(5882):413–417. doi: 10.1038/299413a0. [DOI] [PubMed] [Google Scholar]
  38. Stallcup W. B., Patrick J. Substance P enhances cholinergic receptor desensitization in a clonal nerve cell line. Proc Natl Acad Sci U S A. 1980 Jan;77(1):634–638. doi: 10.1073/pnas.77.1.634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Takai Y., Kikkawa U., Kaibuchi K., Nishizuka Y. Membrane phospholipid metabolism and signal transduction for protein phosphorylation. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1984;18:119–158. [PubMed] [Google Scholar]
  40. Teichberg V. I., Sobel A., Changeux J. P. In vitro phosphorylation of the acetylcholine receptor. Nature. 1977 Jun 9;267(5611):540–542. doi: 10.1038/267540a0. [DOI] [PubMed] [Google Scholar]
  41. Tsien R. W., Giles W., Greengard P. Cyclic AMP mediates the effects of adrenaline on cardiac purkinje fibres. Nat New Biol. 1972 Dec 6;240(101):181–183. doi: 10.1038/newbio240181a0. [DOI] [PubMed] [Google Scholar]
  42. Whiting P. J., Schoepfer R., Swanson L. W., Simmons D. M., Lindstrom J. M. Functional acetylcholine receptor in PC12 cells reacts with a monoclonal antibody to brain nicotinic receptors. Nature. 1987 Jun 11;327(6122):515–518. doi: 10.1038/327515a0. [DOI] [PubMed] [Google Scholar]
  43. Whiting P., Lindstrom J. Purification and characterization of a nicotinic acetylcholine receptor from rat brain. Proc Natl Acad Sci U S A. 1987 Jan;84(2):595–599. doi: 10.1073/pnas.84.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]

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