Skip to main content
SpringerLink
Log in

Cultured chick sympathetic neurons: ATP-induced noradrenaline release and its blockade by nicotinic receptor antagonists

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

The ATP-induced increase in tritium outflow from cultured chick sympathetic neurons prelabelled with [3H]-noradrenaline was investigated.

Seven days-old dissociated cell cultures of embryonic paravertebral ganglia, loaded with [3H]-noradrenaline (0.05 μM), were superfused in the presence of (+)-oxaprotiline and exposed to ATP, ATP-analogues, or 1,1-dimethyl-4-piperazinium (DMPP) for 2 min. ATP (3 μLM-3 mM), 2-methylthio-ATP (3–100 μM), as well as DMPP (10 and 100 μM) induced a significant overflow of tritium. The EC50-value of ATP was 20 μM. Both the ATP-induced and the DMPP-induced tritium overflow was Ca2+-dependent and sensitive to tetrodotoxin (0.3 μM) and ω-conotoxin (0.1 μM); in addition, it was inhibited by the α2-adrenoceptor agonist 5-bromo-6-(2-imidazoline-2-ylamino)-quinoxaline (UK-14,304; 1 μM). The effects of ATP and DMPP were not additive. The ATP-induced as well as the DMPP-induced overflow of tritium was diminished by the P2-purinoceptor antagonists suramin (300 μM) and reactive blue 2 (3 μM); in all 4 cases, the inhibition amouted to approximately 40%. The tritium overflow induced by ATP or DMPP was almost abolished by the nicotinic receptor antagonist mecamylamine (10 μM) and markedly inhibited by hexamethonium (100 μM). Neither ATP nor electrical stimulation caused an overflow of tritium from cultures loaded with [3H]-choline.

The results suggest that ATP at μmolar concentrations induces noradrenaline release from cultured chick sympathetic neurons via an action on a subclass of the nicotinic cholinoceptor.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Akasu T, Koketsu K (1985) Effect of adenosine triphosphate on the sensitivity of the nicotinic acetylcholine-receptor in the bullfrog sympathetic ganglion cell. Br J Pharmacol 84:525–531

    Google Scholar 

  • Allgaier C, Choi KB, Hertting G (1993) Muscarine receptors regulating electrically evoked release of acetylcholine in hippocampus are linked to pertussis toxin-sensitive G-proteins but not to adenylate cyclase. J Neurochem 61:1043–1049

    Google Scholar 

  • Allgaier C, Pullmann F, Schobert A, von Kügelgen I, Heating G (1994a) P2 Purinoceptors modulating noradrenaline release from sympathetic neurons in culture. Eur J Pharmacol 252:R7-R8

    Google Scholar 

  • Allgaier C, Schobert A, Belledin M, Jackisch R, Hertting G (1994b) Modulation of electrically evoked [3H]-noradrenaline release from cultured chick sympathetic neurons. Naunyn-Schmiedeberg's Arch Pharmacol 350:258–266

    Google Scholar 

  • Allgaier C, Wellmann H, Schobert A, von Kügelgen I (1995) Cultured chick sympathetic neurons: modulation of electrically evoked noradrenaline release by P2-purinoceptors. Naunyn-Schmiedeberg's Arch Pharmacol, 352:17–24

    Google Scholar 

  • Ascher P, Large WA, Rang HP (1979) Studies on the mechanism of action of acetylcholine antagonists on rat parasympathetic ganglion cells. J Physiol 295:139–170

    Google Scholar 

  • Boehm S (1994) Noradrenaline release from rat sympathetic neurons evoked by P2-purinoceptor activation. Naunyn-Schmiedeberg's Arch Pharmacol 350:454–458

    Google Scholar 

  • Böhm S, Huck S, Drobny H, Singer EA (1991) Electrically evoked noradrenaline release from cultured chick sympathetic neurons: modulation via presynaptic α2-adrenoceptors and lack of autoinhibition. Naunyn-Schmiedeberg's Arch Pharmacol 344:130–132

    Google Scholar 

  • Bruns RE, Lu GH, Pugsley TA (1986) Characterization of the A2 adenosine receptor labeled by [3H]NECA in rat stritial membranes. Mol Pharmacol 29:331–346

    Google Scholar 

  • Burnstock G, Kennedy C (1985) Is there a basis for distinguishing two types of P2 purinoceptor? Gen Pharmacol 16:433–440

    Article  CAS  PubMed  Google Scholar 

  • Burnstock G, Warland J (1987) P2-Purinoceptors of two subtypes in the rabbit mesenteric artery: reactive blue 2 selectively inhibits responses mediated via the P2y - but not the P2x -purinoceptor. Br J Pharmacol 90: 383–391

    Google Scholar 

  • Cambridge D (1981) UK-14,304, a potent and selective α2-agonist for the characterization of α-adrenoceptor subtypes. Eur J Pharmacol 72:413–415

    Google Scholar 

  • Clones R, Jones S, Brown DA (1993) Zn2+ potentiates ATP-activated currents in rat sympathetic neurons. Pfügers Arch 424:152–158

    Google Scholar 

  • Connolly GP, Harrison PJ, Stone TW (1993) Action of purine and pyrimidine nucleotides on the rat superior cervical ganglion. Br J Pharmacol 110:1297–1304

    Google Scholar 

  • Edwards FA, Gibb AJ (1993) ATP — a fast neurotransmitter. FEBS Lett 325:86–89

    Google Scholar 

  • Edwards FA, Gibb AJ, Coloquhoun D (1992) ATP receptor-mediated synaptic currents in the central nervous system. Nature 359:144–147

    Google Scholar 

  • Evans RJ, Derkarch V, Suprenant A (1992) ATP mediates fast synaptic transmission in mammalian neurons. Nature 357: 503–505

    Google Scholar 

  • Fieber LA, Adams DJ (1991) Adenosine triphosphate-evoked currents in cultured neurones dissociated from rat parasympathetic cardiac ganglia. J Physiol 434:239–256

    Google Scholar 

  • Greene LA, Rein G (1978) Release of norepinephrine from neurons in dissociated cell cultures of chick sympathetic ganglia via stimulation of nicotinic and muscarinic acetylcholine receptors. J Neurochem 30:579–586

    Google Scholar 

  • Igusa Y (1988) Adenosine 5′-triphosphate activates acetylcholine receptor channels in cultured Xenopus myotomal muscle cells. J Physiol 405:169–185

    Google Scholar 

  • Illes P, Nörenberg W (1993) Neuronal ATP receptors and their mechanism of action. Trends Pharmacol Sci 14:50–54

    Google Scholar 

  • Inoue K, Nakazawa K, Fujimori K, Tanaka A (1989) Extracellular adenosine 5′-triphosphate-evoked norepinephrine secretion not relating to voltage-gated Ca channels in pheochromocytoma PC12 cells. Neurosci Lett 106:294–299

    Google Scholar 

  • Inoue K; Nakazawa K, Fujimori K, Watano T, Tanaka A (1992) Extracellular adenosine 5′-triphosphate-evoked glutamate release in cultured hippocampal neurons. Neurosci Lett 134:215–218

    Google Scholar 

  • Jahr CE, Jessell TM (1983) ATP excites a subpopulation of rat dorsal horn neurones. Nature 304:730–733

    Google Scholar 

  • Katayama Y, Morita K (1989) Adenosine 5′-triphosphate modulates membrane potassium conductance in guinea-pig myenteric neurones. J Physiol 408:373–390

    Google Scholar 

  • Kennedy C (1990) P1- and P2-purinoceptor subtypes — an update. Arch Int Pharmacodyn 303:30–50

    Google Scholar 

  • Kim KT, Westhead EW (1989) Cellular responses to Ca2+ from extracellular and intracellular sources are different as shown by simultaneous measurements of cytosolic Ca2+ and secretion from bovine chromaffin cells. Proc Natl Acad Sci USA 86:9881–9885

    Google Scholar 

  • Löffelholz K (1979) Release induced by nicotinic agonists. In: Paton DM (ed) The release of catecholamines from adrenergic neurons. Pergamon Press, Oxford, pp 275–301

    Google Scholar 

  • Moody C, Burnstock G (1982) Evidence of the presence of P1-purinoceptors on cholinergic nerve terminals in the guinea pig ileum. Eur J Pharmacol 77:1–9

    Google Scholar 

  • Nakazawa K (1994) ATP-activated current and its interaction with acetylcholine-activated current in rat sympathetic neurons. J Neurosci 14:740–750

    Google Scholar 

  • Nakazawa K, Inoue K (1993) ATP- and acetylcholine-activated channels co-existing in cell-free membrane patches from rat sympathetic neuron. Neurosci Lett 163:97–100

    Google Scholar 

  • Phillis JW, Kostopoulos GK, Limacher JJ (1975) A potent depressant action of adenine derivatives on cerebral cortical neurones. Eur J Pharmacol 30:125–129

    Google Scholar 

  • Reekie FM, Burnstock G (1994) Some effects of purines on neurones of guinea-pig superior cervical ganglia. Gen Pharmacol 25: 143–148

    Google Scholar 

  • Salt TE, Hill RG (1983) Excitation of single sensory neurones in the rat caudal trigeminal nucleus by iontophoretically applied adenosine 5′-triphosphate. Neurosci Lett 35:53–57

    Google Scholar 

  • Shen K-Z, North RA (1993) Excitation of rat locus coeruleus neurons by adenosine 5′-triphosphate: ionic mechanism and receptor characterization. J Neurosci 13:894–899

    Google Scholar 

  • Silinsky EM, Gerzanich V, Vanner SM (1992) ATP mediates excitatory synaptic transmission in mammalian neurones. Br J Pharmacol 106:762–763

    Google Scholar 

  • Starke K (1977) Regulation of noradrenaline release by presynaptic receptor systems. Rev Physiol Biochem Pharmacol 77:1–124.

    Google Scholar 

  • Tschöpl M, Harms L, Nörenberg W, Illes P (1992) Excitatory effects of adenosine 5′-triphosphate on rat locus coeruleus neurones. Eur J Pharmacol 213:71–77

    Google Scholar 

  • von Kügelgen I, Schöffel E, Starke K (1989) Inhibition by nucleotides acting at presynaptic P2-receptors of sympathetic neuroeffector transmission in the mouse isolated vas deferens. Naunyn-Schmiedeberg's Arch Pharmacol 340:522–532

    Google Scholar 

  • Varanda WA, Aracava Y, Sherby SM, van Meter WG, Eldefrawi ME, Albuquerque EX (1985) The acetylcholine receptor of the neuromuscular junction recognizes mecamylamine as a noncompetitive antagonist. Mol Pharmacol 28:128–137

    Google Scholar 

  • Wakade AR, Wakade TD (1988) Comparison of transmitter release properties of embryonic sympathetic neurons growing in vivo and in vitro. Neuroscience 27:1007–1019

    Google Scholar 

  • Waldmeier PC, Baumann PA, Hauser K, Maitre L, Storm A (1982) Oxaprotiline, a noradrenaline uptake inhibitor with an active and an inactive enantiomer. Biochem Pharmacol 31:2169–2176.

    Google Scholar 

  • Zimmermann H (1994) Signalling via ATP in the nervous system. Trends Neurosci 17:420–426

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Pharmacology and Toxicology, Albert-Ludwigs University of Freiburg, Hermann-Herder-Strasse 5, D-79104, Freiburg, Germany

    Clemens Allgaier, Henning Wellmann, Angelika Schobert & Ivar von Kügelgen

  2. Department of Biochemistry, Chemical Laboratories, Albertstrasse 21, D-79104, Freiburg, Germany

    Gerhart Kurz

Authors
  1. Clemens Allgaier
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Henning Wellmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Angelika Schobert
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gerhart Kurz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ivar von Kügelgen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Allgaier, C., Wellmann, H., Schobert, A. et al. Cultured chick sympathetic neurons: ATP-induced noradrenaline release and its blockade by nicotinic receptor antagonists. Naunyn-Schmiedeberg's Arch Pharmacol 352, 25–30 (1995). https://doi.org/10.1007/BF00169186

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00169186

Key words

Search

Navigation