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Tyrosine hydroxylase activation and transmitter release from central noradrenergic neurons by electrical field stimulation

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Summary

Electrical stimulation of the noradrenergic neurons in the locus coeruleus of the rat results in a marked increase in the tyrosine hydroxylase activity of the hippocampus on the stimulated side (Roth et al., 1975). We have developed an in vitro system to further study this interesting phenomenon. Rat hippocampal slices were stimulated in an electrical field using specially built chambers which permitted a continous superfusion with Krebs Ringer Phosphate solution while stimulation was taking place. The slices were then homogenized and tyrosine hydroxylase activity and kinetic parameters were determined in the 104000 g supernate fraction. Electrical field stimulation (60 V, 4 ms, 5–20 Hz, 5 min) induced a stimulus-dependent increase in tyrosine hydroxylase activity. The increase in tyrosine hydroxylase activity was directly dependent on the number of pulses of stimulation applied. Potassium depolarization of hippocampal slices also resulted in a marked increase in the activity of the enzyme. Electrical field stimulation appears to activate tyrosine hydroxylase by causing an increase in its affinity for both substrate and pteridine cofactor and by decreasing its affinity for the endproduct inhibitor, norepinephrine. No change in Vmax was observed. The superfusion system was also used to study spontaneous and electrically evoked release of labelled norepinephrine from hippocampal slices. Electrical field stimulation for 1.0 min produced a marked increase in the release of label. Absence of calcium from the superfusion solution almost completely abolished the electrically evoked release of exogenously taken up 3H-norepinephrine. The release of 3H-norepinephrine was also found to be dependent on the number of pulses of stimulation used. The results reported support the hypothesis that depolarization of central noradrenergic neurons results in an increase in transmitter synthesis mediated in part via a kinetic activation of tyrosine hydroxylase. The concomitant use of superfusion and electrical field stimulation of hippocampal slices provides a simple system to study neurotransmitter synthesis and release in central noradrenergic neurons.

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References

  • 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.) 56, 1491–1496 (1966)

    Google Scholar 

  • Baldessarini, R. J., Kopin, I. J.: The effect of drugs on the release of 3H-norepinephrine from central nervous system tissues by electrical stimulation in vitro. J. Pharmacol. exp. Ther. 156, 31–38 (1967)

    Google Scholar 

  • Baker, P. F.: Transport and metabolism of calcium ions in nerve. Prog. Biophys. Mol. Biol. 24, 177–223 (1972)

    Google Scholar 

  • Boullin, D. J.: The action of extracellular cations on the release of the sympathetic transmitter from peripheral nerves. J. Physiol. (Lond.) 189, 85–99 (1967)

    Google Scholar 

  • Burn, J. H., Gibbons, W. R.: The part played by calcium in determining the response to stimulation of sympathetic postganglionic fibres. Br. J. Pharmacol. Chemother. 22, 540–548 (1964)

    Google Scholar 

  • Bustos, G., Roth, R. H.: Release of monoamines from the striatum and hypothalamus: effect of γ-hydroxybutyrate. Br. J. Pharmacol. 46, 101–115 (1972)

    Google Scholar 

  • Bustos, G., Roth, R. H., Morgenroth, V. I., III: Activation of tyrosine hydroxylase in rat striatal slices by K+-depolarization — effect of ethanol. Biochem. Pharmacol. 25, 2493–2497 (1976)

    Google Scholar 

  • Coyle, J. T.: Tyrosine hydroxylase in rat brain — cofactor requirements, regional and subcellular distribution. Biochem. Pharmacol. 21, 1935–1944 (1972)

    Google Scholar 

  • Dixon, M.: The determination of enzyme inhibitor constants. Biochem. J. 55, 170–171 (1953)

    Google Scholar 

  • Douglas, W. W.: The mechanism of release of catecholamines from the adrenal medulla. Pharmacol. Rev. 18, 471–480 (1966)

    Google Scholar 

  • Gordon, R., Reid, J. V. D., Sjoerdsma, A., Udenfriend, S.: Increased synthesis of norepinephrine in the rat heart on electrical stimulation of the stellate ganglion. Mol. Pharmacol. 6, 606–613 (1966)

    Google Scholar 

  • Gutman, Y., Segal, J.: Effect of calcium, potassium and sodium on tyrosine hydroxylase activity in different regions of rat brain. Biochem. Pharmacol. 22, 865–868 (1973)

    Google Scholar 

  • Harris, J. E., Roth, R. H.: Potassium-induced acceleration of catecholamine biosynthesis in brain slices. A study on the mechanism of action. Mol. Pharmacol. 7, 593–604 (1971)

    Google Scholar 

  • Kakiuchi, S., Rall, T. W., McIlwain, H.: The effect of electrical stimulation upon the accumulation of adenosine 3′,5′-phosphate in isolated cerebral tissue. J. Neurochem. 16, 485–491 (1969)

    Google Scholar 

  • Katz, I. R., Kopin, I. J.: Electrical field-stimulated release of 3H-norepinephrine from rat atrium. J. Pharmacol. Exp. Ther. 169, 229–236 (1969a)

    Google Scholar 

  • Katz, I. R., Kopin, I. J.: Release of 3H-norepinephrine and 3H-serotonin from brain slices by electrical-field stimulation. Calcium dependence and the effects of lithium, ouabain, and tetrodoxin. Biochem. Pharmacol. 18, 1935–1939 (1969b)

    Google Scholar 

  • Kaufman, S.: Phenylalanine hydroxylase. Meth. Enzym. 5, 809–816 (1962)

    Google Scholar 

  • Korf, J., Aghajanian, G. K., Roth, R. H.: Stimulation and destruction of the locus coeruleus: opposite effects on 3-methoxy-4-hydroxyphenylglycol sulfate levels in the rat cerebral cortex. Eur. J. Pharmacol. 21, 305–310 (1973a)

    Google Scholar 

  • Korf, J., Roth, R. H., Aghajanian, G. K.: Alterations in turnover and endogenous levels of norepinephrine in the cerebral cortex following electrical stimulation and acute axotomy of cerebral noradrenergic pathways. Eur. J. Pharmacol. 23, 276–282 (1973b)

    Google Scholar 

  • Kuczenski, R. T., Mandell, A. S.: Allosteric activation of hypothalamic tyrosine hydroxylase by ions and sulphated mucopolysaccharides. J. Neurochem. 19, 131–137 (1972)

    Google Scholar 

  • Lerner, P., Ames, M. M., Lovenberg, W.: The effect of ethylene glycol bis(β-aminoethyl ether)-N′,N′-tetraacetic acid and calcium on tyrosine hydroxylase activity. Mol. Pharmacol. 13, 44–49 (1977)

    Google Scholar 

  • Lineweaver, H., Burk, D.: The determination of enzyme dissociation constants. J. Amer. Chem. Soc. 56, 658–666 (1934)

    Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurements with the folin reagent. J. Biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Morgenroth, V. H., III, Boadle-Biber, M. C., Roth, R. H.: Tyrosine hydroxylase: Activation by nerve stimulation. Proc. Natl. Acad. Sci. (U.S.A.) 71, 4283–4287 (1974)

    Google Scholar 

  • Morgenroth, V. H., III, Boadle-Biber, M. C., Roth, R. H.: Activation of tyrosine hydroxylase from central noradrenergic neurons by calcium. Mol. Pharmacol. 11, 427–435 (1975)

    Google Scholar 

  • Morgenroth, V. H., III, Hegstrand, L. R., Roth, R. H., Greengard, P.: Evidence for involvement of protein kinase in the activation by cAMP of brain tyrosine-3-monooxygenase. J. Biol. Chem. 250, 1946–1948 (1975)

    Google Scholar 

  • Osborne, N. N., Neuhoff, V.: Activation of snail (Helix pomatia) nervous tissue tyrosine monooxygenase by calcium in vitro. Hoppe-Seylers Z. Physiol. Chem. 357, 1271–1275 (1976)

    Google Scholar 

  • Roth, R. H., Stjärne, L., von Euler, U. S.: Acceleration of noradrenaline biosynthesis by nerve stimulation. Life Sci. 5, 1071–1075 (1966)

    Google Scholar 

  • Roth, R. H., Stjärne, L., von Euler, U. S.: Factors influencing the rate of norepinephrine biosynthesis in nerve tissue. J. Pharmacol. Exp. Ther. 158, 373–377 (1967)

    Google Scholar 

  • Roth, R. H., Morgenroth, V. H., III, Salzman, P. M.: Tyrosine hydroxylase: Allosteric activation induced by stimulation of central noradrenergic neurons. Naunyn-Schmiedeberg's Arch. Pharmacol. 289, 327–343 (1975)

    Google Scholar 

  • Salzman, P. M., Roth, R. H.: Noradrenergic neurons: Post-stimulation increase in catecholamine biosynthesis. Proc. 10th CINP Congress. Oxford-New York: Pergamon Press (in press), 1978

    Google Scholar 

  • Sedvall, G. C., Kopin, I. G.: Acceleration of norepinephrine synthesis in the rat submaxillary gland in vivo during sympathetic nerve stimulation. Life Sci. 6, 45–51 (1967)

    Google Scholar 

  • Shiman, R., Akino, M., Kaufman, S.: Solubilization and partial purification of tyrosine hydroxylase from bovine adrenal medulla. J. Biol. Chem. 246, 1330–1340 (1971)

    Google Scholar 

  • Shimizu, H., Creveling, C. R., Daly, J. W.: 3′,5′-Cyclic adenosine monophosphate formation in brain slices; stimulation by Batrachotoxin, ouabain, veratridine, and potassium ions. Mol. Pharmacol. 6, 184–198 (1970)

    Google Scholar 

  • Weiner, N.: Regulation of norepinephrine biosynthesis. Ann. Rev. Pharmacol. 10, 273–293 (1970)

    Google Scholar 

  • Weiner, N., Lee, F.-L.: The role of calcium in norepinephrine release and synthesis. In: Chemical Tools in Catecholamine Research II (Almgren, Carlsson, Engel, eds.). New York: Amer. Elsevier Publ. Co., 1975

    Google Scholar 

  • Wilkinson, G. N.: Statistical estimations in enzyme kinetics. Biochem. J. 80, 324–332 (1961)

    Google Scholar 

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Authors and Affiliations

  1. Department of Neurobiology, Institute of Biological Sciences, Catholic University, Santiago, Chile

    Gonzalo Bustos & Juan Luis Hancke

  2. Departments of Pharmacology and Psychiatry, Yale University School of Medicine, 06510, New Haven, Connecticut, USA

    Robert H. Roth

Authors
  1. Gonzalo Bustos
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  2. Robert H. Roth
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  3. Victor H. Morgenroth III
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  4. Juan Luis Hancke
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Additional information

A preliminary account of this work was reported at the eight Meeting of the Argentinian Society for Experimental Pharmacology held November 14–18, 1975, at San Luis, Argentina

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Bustos, G., Roth, R.H., Morgenroth, V.H. et al. Tyrosine hydroxylase activation and transmitter release from central noradrenergic neurons by electrical field stimulation. Naunyn-Schmiedeberg's Arch. Pharmacol. 301, 149–156 (1978). https://doi.org/10.1007/BF00507030

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  • DOI: https://doi.org/10.1007/BF00507030

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