Summary
In mouse atria previously incubated with [3H]-noradrenaline, carbachol (1.0 µmol/l) significantly inhibited the fractional stimulation-induced (S-I) outflow of radioactivity. The inhibitory effect of carbachol was greater in the presence of the α-adrenoceptor antagonist phentolamine (1.0 µmol/l), which by itself significantly increased the S-I outflow of radioactivity. In both cases the inhibitory effect of carbachol was blocked by atropine (0.3 µmol/l), suggesting that the effect was mediated through muscarinic receptors. 8-Bromo cyclic AMP (270 µmol/l) in the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX,100 µmol/l), was used to maximally enhance the S-I outflow of radioactivity through the cyclic AMP mechanism. The inhibitory effect of carbachol either in the presence or in the absence of phentolamine, was not reduced in the presence of 8-bromo cyclic AMP and IBMX. Similar results with carbachol in the presence of 8-bromo cyclic AMP and IBMX were also found in rat right atrial strips which had been incubated with [3H]-noradrenaline. These results suggest that the effects through inhibitory prejunctional muscarinic receptors are not mediated by cyclic AMP.
The protein kinase inhibitor, staurosporine (0.1 µmol/l), significantly blocked the enhancing effects of 8-bromo cyclic AMP (270 µmol/l) plus IBMX (100 µmol/l) on the S-I outflow of radioactivity from rat atrial strips. The inhibitory effect of carbachol (1.0 µmol/l) however, was not reduced in the presence of staurosporine, suggesting that protein kinases affected by staurosporine (protein kinase A, protein kinase C) are not involved in the postreceptor mechanism for inhibitory prejunctional muscarinic receptors. This finding further rules out the involvement of cyclic AMP in muscarinic inhibition.
The inhibitory effect of carbachol either by itself or in the presence of phentolamine, was not reduced in atria from mice that had been pretreated with pertussis toxin (1.5 or 3.0 µg). Furthermore, in rat atrial strips, the inhibitory effect of carbachol either in the presence or in the absence of phentolamine, was also not altered by pretreating the rats with pertussis toxin (8.4 µg). The results suggest that in both tissues the major mechanism for inhibition of noradrenaline release through muscarinic receptors does not involve a pertussis toxin sensitive G protein.
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References
Beavo JA (1988) Multiple isoenzymes of cyclic nucleotide phosphodiesterase. Adv Second Messenger Phosphoprotein Res 22:1–33
Beebe SJ, Beasley-Leach A, Corbin JD (1988) cAMP analogues used to study low-Km hormone-sensitive phosphodiesterase. Methods Enzymol 159:531–540
Daly JW (1983) Adenosine receptors: characterization with radioactive ligands. In: Daly JW, Kuroda Y, Phillis JW, Shimizu H, Ui M (eds) Physiology and pharmacology of adenosine derivatives. Raven, New York, pp 59–69
Endoh M, Maruyama M, Iijima T (1985) Attenuation of muscarinic cholinergic inhibition by islet-activating protein in the heart. Am J Physiol 249:H309-H320
Enero MA, Saidman BQ (1977) Possible feedback inhibition of noradrenaline release by purine compounds. Naunyn-Schmiedeberg's Arch Pharmacol 297:39–46
Fredholm BB, Brodnik K, Strandberg K (1979) On the mechanism of relaxation of tracheal muscle by theophylline and cyclic nucleotide phosphodiesterase inhibitors. Acta Pharmacol Toxicol 45:336–344
Hedqvist P (1974) Prostaglandin action on noradrenaline release and mechanical responses in the stimulated guinea pig vas deferens. Acta Physiol Scand 90:86–93
Johnston H, Majewski H, Musgrave IF (1987) Involvement of cyclic nucleotides in prejunctional modulation of noradrenaline release in mouse atria. Br J Pharmacol 91:773–781
Kalsner S, Quillan M, (1988) Presynaptic interactions between acetylcholine and adrenergic antagonists on norepinephrine release. J Pharmacol Exp Ther 244:879–891
Kurose H, Ui M, (1983) Functional uncoupling of muscarinic receptors from adenylate cyclase in rat cardiac membranes by the active component of islet-activating protein, pertussis toxin. J Cyclic Nucleotide Protein Phosphor Res 9(4–5):305–318
Levitski A (1986) β-Adrenergic receptors and their mode of coupling to adenylate cyclase. Physiol Rev 66:819–854
Levy MN, Blattberg B (1976) Effect of vagal stimulation on the overflow of norepinephrine into the coronary sinus during cardiac sympathetic nerve stimulation in the dog. Circ Res 38:81–85
Limberger N, Späth L (1987) Interaction between presynaptic receptors at the noradrenergic axons of rabbit brain cortex. Naunyn-Schmiedeberg's Arch Pharmacol 335: R84
Limberger N, Späth L, Hölting Th, Starke K (1986) Mutual interaction between presynaptic α2-adrenoceptors and opioid k-receptors at the noradrenergic axons of rabbit brain cortex. Naunyn-Schmiedeberg's Arch Pharmacol 334:166–171
Loiacono RE, Rand MJ, Story DF (1985) Interaction between the inhibitory action of acetylcholine and the α-adrenoceptor autoinhibitory feedback system on release of [3H]-noradrenaline from rat atria and rabbit ear artery. Br J Pharmacol 84:697–705
Majewski H, Ishac EJN, Musgrave IF (1987) Intraneuronal mechanisms involved in the modulation of noradrenaline release through prejunctional receptors. In: Bevan JA, Maxwell RA, Majewski H, Story DF (eds) Vascular Neuroeffector Mechanisms IRL, Oxford Washington, pp 243–250
Marley PB, Wilson JV, Habel H, Blaskett A (1985) Specific anti-Bordetella pertussis activities in human blood examined by enzyme-linked immunoassay and biological assay. Dev Biol Stand 61:341–352
Mei L, Yamamura HI, Roeske WR (1988) Muscarinic receptor-mediated hydrolysis of phosphatidylinositols in human neuroblastoma (SH-SY5Y) cells is sensitive to pertussis toxin. Brain Res 447:360–363
Molderings G (1989) Interaction between presynaptic α2-adrenoceptor and 5-HT1B receptor on the sympathetic nerve terminals of the rat vena cava. Naunyn-Schmiedeberg's Arch Pharmacol 339 (suppl):R95
Muscholl E (1980) Peripheral muscarinic control of norepinephrine release in the cardiovascular system. Am J Physiol 239:H713-H720
Musgrave IF, Marley P, Majewski H (1987) Pertussis toxin does not attenuate α2-adrenoceptor mediated inhibition of noradrenaline release in mouse atria. Naunyn-Schmiedeberg's Arch Pharmacol 336:280–286
Nathanson NM (1987) Molecular properties of the muscarinic acetylcholine receptor. Annu Rev Neurosci 10:195–236
Ramme D, Illes P, Späth L, Starke K (1986) Blockade of α2-adrenoceptors permits the operation of otherwise silent opioid k-receptors at the sympathetic axons of rabbit jejunal arteries. Naunyn-Schmiedeberg's Arch Pharmacol 334:48–55
Rand MJ, Majewski H, Wong-Dusting H, Story DF, Loiacono RE, Ziogas J (1987) Modulation of neuroeffector transmission. J Cardiovasc Pharmacol 10 (Suppl. 12):S33-S44
Rüegg UT, Burgess GM (1989) Staurosporine, K-252 and UCN-01: potent but nonspecific inhibitors of protein kinases. Trends Pharmacol Sci 10:218–220
Schächtele C, Rudolph C, Osswald H (1989) Inhibition of protein kinase (PKC) and three other protein kinases by several reference CKC inhibitors. Naunyn-Schmiedeberg's Arch Pharmacol 340 (suppl):R36
Tamaoki T, Nomoto H, Takahashi I, Kato Y, Morimoto M, Tomito F (1986) Staurosporine, a potent inhibitor of phospholipid/Ca2+ dependent protein kinase. Biochem Biophys Res Commun 135:397–402
Vanhoutte PM, Levy MN (1980) Prejunctional cholinergic modulation of adrenergic neurotransmission in the cardiovascular system. Am J Physiol 238:H275-H281
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Costa, M., Majewski, H. Inhibitory prejunctional muscarinic receptors at sympathetic nerves do not operate through a cyclic AMP dependent pathway. Naunyn-Schmiedeberg's Arch Pharmacol 342, 630–639 (1990). https://doi.org/10.1007/BF00175705
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DOI: https://doi.org/10.1007/BF00175705