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. 1984 Nov;81(22):7243–7247. doi: 10.1073/pnas.81.22.7243

Electrical stimulation increases phosphorylation of tyrosine hydroxylase in superior cervical ganglion of rat.

A L Cahill, R L Perlman
PMCID: PMC392115  PMID: 6150485

Abstract

Electrical stimulation of the superior cervical ganglion of the rat increased the phosphorylation of tyrosine hydroxylase (tyrosine 3-monooxygenase, EC 1.14.16.2) in this tissue. Ganglia were incubated with [32P]Pi for 90 min and were then electrically stimulated via the preganglionic nerve. Tyrosine hydroxylase was isolated from homogenates of the ganglia by immunoprecipitation followed by polyacrylamide gel electrophoresis. 32P-labeled tyrosine hydroxylase was visualized by radioautography, and the incorporation of 32P into the enzyme was quantitated by densitometry of the radioautograms. Stimulation of ganglia at 20 Hz for 5 min increased the incorporation of 32P into tyrosine hydroxylase to a level 5-fold that found in unstimulated control ganglia. The increase in phosphorylation of tyrosine hydroxylase was dependent on the duration and frequency of stimulation. Preganglionic stimulation did not increase the phosphorylation of tyrosine hydroxylase in a medium that contained low Ca2+ and high Mg2+. Increases in phosphorylation were reversible; within 30 min after the cessation of stimulation, the incorporation of 32P into tyrosine hydroxylase decreased to the level found in unstimulated ganglia. The nicotinic antagonist hexamethonium reduced the increase in 32P incorporation into tyrosine hydroxylase by about 50%, while the muscarinic antagonist atropine had no effect. Thus, preganglionic stimulation appeared to increase the phosphorylation of tyrosine hydroxylase in part by a nicotinic mechanism and in part by a noncholinergic mechanism. Antidromic stimulation of ganglia also increased the phosphorylation of tyrosine hydroxylase. Two-dimensional gel electrophoresis revealed that electrical stimulation also increased the incorporation of 32P into at least six other phosphoproteins in the ganglion.

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Selected References

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  1. Alousi A., Weiner N. The regulation of norepinephrine synthesis in sympathetic nerves: effect of nerve stimulation, cocaine, and catecholamine-releasing agents. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1491–1496. doi: 10.1073/pnas.56.5.1491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrews D. W., Langan T. A., Weiner N. Evidence for the involvement of a cyclic AMP-independent protein kinase in the activation of soluble tyrosine hydroxylase from rat striatum. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2097–2101. doi: 10.1073/pnas.80.8.2097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Greengard P. Intracellular signals in the brain. Harvey Lect. 1979 1980;75:277–331. [PubMed] [Google Scholar]
  4. Haycock J. W., Meligeni J. A., Bennett W. F., Waymire J. C. Phosphorylation and activation of tyrosine hydroxylase mediate the acetylcholine-induced increase in catecholamine biosynthesis in adrenal chromaffin cells. J Biol Chem. 1982 Nov 10;257(21):12641–12648. [PubMed] [Google Scholar]
  5. Horwitz J., Perlman R. L. Stimulation of DOPA synthesis in the superior cervical ganglion by veratridine. J Neurochem. 1984 Feb;42(2):384–389. doi: 10.1111/j.1471-4159.1984.tb02689.x. [DOI] [PubMed] [Google Scholar]
  6. Ikeno T., Dickens G., Lloyd T., Guroff G. The receptor-mediated activation of tyrosine hydroxylation in the superior cervical ganglion of the rat. J Neurochem. 1981 May;36(5):1632–1640. doi: 10.1111/j.1471-4159.1981.tb00413.x. [DOI] [PubMed] [Google Scholar]
  7. Ip N. Y., Ho C. K., Zigmond R. E. Secretin and vasoactive intestinal peptide acutely increase tyrosine 3-monooxygenase in the rat superior cervical ganglion. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7566–7569. doi: 10.1073/pnas.79.23.7566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ip N. Y., Perlman R. L., Zigmond R. E. Acute transsynaptic regulation of tyrosine 3-monooxygenase activity in the rat superior cervical ganglion: evidence for both cholinergic and noncholinergic mechanisms. Proc Natl Acad Sci U S A. 1983 Apr;80(7):2081–2085. doi: 10.1073/pnas.80.7.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ip N. Y., Perlman R. L., Zigmond R. E. Both nicotinic and muscarinic agonists acutely increase tyrosine 3-monooxygenase activity in the superior cervical ganglion. J Pharmacol Exp Ther. 1982 Nov;223(2):280–283. [PubMed] [Google Scholar]
  10. Joh T. H., Park D. H., Reis D. J. Direct phosphorylation of brain tyrosine hydroxylase by cyclic AMP-dependent protein kinase: mechanism of enzyme activation. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4744–4748. doi: 10.1073/pnas.75.10.4744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Letendre C. H., MacDonnell P. C., Guroff G. The biosynthesis of phosphorylated tyrosine hydroxylase by organ cultures of rat medulla and superior cervical ganglia. Biochem Biophys Res Commun. 1977 Feb 7;74(3):891–897. doi: 10.1016/0006-291x(77)91602-3. [DOI] [PubMed] [Google Scholar]
  13. Meligeni J. A., Haycock J. W., Bennett W. F., Waymire J. C. Phosphorylation and activation of tyrosine hydroxylase mediate the cAMP-induced increase in catecholamine biosynthesis in adrenal chromaffin cells. J Biol Chem. 1982 Nov 10;257(21):12632–12640. [PubMed] [Google Scholar]
  14. Morgenroth V. H., 3rd, Boadle-Biber M., Roth R. H. Tyrosine hydroxylase: activation by nerve stimulation. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4283–4287. doi: 10.1073/pnas.71.11.4283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nestler E. J., Greengard P. Nerve impulses increase the phosphorylation state of protein I in rabbit superior cervical ganglion. Nature. 1982 Apr 1;296(5856):452–454. doi: 10.1038/296452a0. [DOI] [PubMed] [Google Scholar]
  16. Noon J. P., McAfee D. A., Roth R. H. Norepinephrine release from nerve terminals within the rabbit superior cervical ganglion. Naunyn Schmiedebergs Arch Pharmacol. 1975;291(2):139–162. doi: 10.1007/BF00500046. [DOI] [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Raese J. D., Edelman A. M., Makk G., Bruckwick E. A., Lovenberg W., Barchas J. D. Brain striatal tyrosine hydroxylase: activation of the enzyme by cyclic AMP-independent phosphorylation. Commun Psychopharmacol. 1979;3(5):295–301. [PubMed] [Google Scholar]
  19. Roth R. H., Morgenroth V. H., 3rd, Salzman P. M. Tyrosine hydroxylase: allosteric activation induced by stimulation of central noradrenergic neurons. Naunyn Schmiedebergs Arch Pharmacol. 1975;289(4):327–343. doi: 10.1007/BF00508408. [DOI] [PubMed] [Google Scholar]
  20. Roth R. H., Salzman P. M., Morgenroth V. H., 3rd Noradrenergic neurons: allosteric activation of hippocampal tyrosine hydroxylase by stimulation of the locus coeruleus. Biochem Pharmacol. 1974 Oct 1;23(19):2779–2784. doi: 10.1016/0006-2952(74)90051-3. [DOI] [PubMed] [Google Scholar]
  21. Steinberg M. I., Keller C. E. Enhanced catecholamine synthesis in isolated rat superior cervical ganglia caused by nerve stimulation: dissociation between ganglionic transmission and catecholamine synthesis. J Pharmacol Exp Ther. 1978 Feb;204(2):384–399. [PubMed] [Google Scholar]
  22. Stephens J. K., Masserano J. M., Vulliet P. R., Weiner N., Nakane P. K. Immunocytochemical localization of tyrosine hydroxylase in rat adrenal medulla by the peroxidase labeled antibody method: effects of enzyme activation on ultrastructural distribution of the enzyme. Brain Res. 1981 Mar 30;209(2):339–354. doi: 10.1016/0006-8993(81)90158-x. [DOI] [PubMed] [Google Scholar]
  23. Volle R. L., Patterson B. A. Regulation of cyclic AMP accumulation in a rat sympathetic ganglion: effects of vasoactive intestinal polypeptide. J Neurochem. 1982 Oct;39(4):1195–1197. doi: 10.1111/j.1471-4159.1982.tb11516.x. [DOI] [PubMed] [Google Scholar]
  24. Volle R. L., Quenzer L. F., Patterson B. A. The regulation of cyclic nucleotides in a sympathetic ganglion. J Auton Nerv Syst. 1982 Jul;6(1):65–72. doi: 10.1016/0165-1838(82)90023-6. [DOI] [PubMed] [Google Scholar]
  25. Weiner N., Lee F. L., Dreyer E., Barnes E. The activation of tyrosine hydroxylase in noradrenergic neurons during acute nerve stimulation. Life Sci. 1978 Apr 3;22(13-15):1197–1215. doi: 10.1016/0024-3205(78)90088-7. [DOI] [PubMed] [Google Scholar]
  26. Weiner N. Regulation of norepinephrine biosynthesis. Annu Rev Pharmacol. 1970;10:273–290. doi: 10.1146/annurev.pa.10.040170.001421. [DOI] [PubMed] [Google Scholar]
  27. Yamauchi T., Fujisawa H. In vitro phosphorylation of bovine adrenal tyrosine hydroxylase by adenosine 3':5'-monophosphate-dependent protein kinase. J Biol Chem. 1979 Jan 25;254(2):503–507. [PubMed] [Google Scholar]
  28. Yamauchi T., Fujisawa H. Regulation of bovine adrenal tyrosine 3-monooxygenase by phosphorylation-dephosphorylation reaction, catalyzed by adenosine 3':5'-monophosphate-dependent protein kinase and phosphoprotein phosphatase. J Biol Chem. 1979 Jul 25;254(14):6408–6413. [PubMed] [Google Scholar]

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