Abstract
Catecholamine secretion in the bovine adrenal medulla is evoked largely by nicotinic receptor activation. However, bovine adrenal medulla also contain muscarinic receptors that mediate several cell responses. To understand the physiological role of muscarinic receptors in the bovine adrenal medulla it is important to identify the pharmacological subtypes present in this tissue. For this, we analyzed the abilities of differnt selective muscarinic antagonists in displacing the binding of the non-selective antagonist [3H] quinuclidinyl benzylate to an enriched plasma membrane fraction prepared from bovine adrenal medulla. All the selective antagonists bind at least two bindings sites with different affinities. The binding profile of the sites with high proportion is similar to the M2 subtype and those present in low proportion have a M1 profile. However, some variation in the proportion of the sites for the different ligands suggest the presence of the third pharmacological subtype (M3). We conclude that the sites in high proportion (60–80%) correspond to M2 muscarinic subtypes, and the rest is constitute by M1 plus M3 subtypes. The presence of multiplicity of subtypes in the adrenal medulla membranes suggests a diversity of functions of muscarinic receptors in the adrenal gland.
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Abbreviations
- [3H]QNB:
-
[3H]quinuclidinyl benzylate
- HHSiD:
-
hexahydro-siladifenidol-hydrochloride
- AF-DX 116:
-
11-[[2-(diethylamino)methyl]]-1-piperidinyl]-5,11-dihydro-6H-pyrido[2,3,-b][1,4]benzodiazepin-6-one
- 4-DAMP:
-
4-diphenylacetoxy-N-methyl piperidine methobromide
References
Ledbetter, F. H., and Kirshner, N. 1975. Studies of chick adrenal medulla in organ culture. Biochem. Pharmacol. 24:967–974.
Knight, D. E., and Baker, P. F. 1986. Observation of the muscarinic receptors of bovine adrenal medulla. Neuroscience 19:357–366.
Ballesta, J. J., Borges, R., García, A. G., and Hidalgo, M. J. 1989. Secretory and radioligand binding studies on muscarinic receptors in bovine and feline chromaffin cell. J. Physiol. 418:411–426.
Liang, B. T., and Perlman, R. L. 1979. Catecholamine secretion by hamster adrenal cells. J. Neurochem. 32:927–933.
Almazán, G., Aunis, D., García, A. G., Montiel, C., Nicolás, G. P., and Sánchez-García, P. 1984. Effects of collagenase on the release of 3H-noradrenaline from culture bovine adrenal chromaffin cells. Br. J. Pharmacol. 81:599–610.
Barron, B. A., Murrin, L. C., and Hexum, T. D. 1986. Muscarinic binding sites in bovine adrenal medulla. Eur. J. Pharmacol. 122:269–273.
Kayaalp, S. O., and Neff, N. H. 1979. Cholinergic muscarinic receptors of bovine adrenal medulla. Neuropharmacology 18:909–911.
Kao, L.-S., and Howthorne, J. N. 1985. Calcium mobilization and catecholamine secretion in adrenal chromaffin cells. J. Biol. Chem. 260:2019–2022.
Derome, G., Tseng, R., Mercier, P., Lemaire, J., and Lemaire, S. 1981. Possible muscarinic regulation of catecholamine secretion mediated by cyclic GMP in isolated bovine chromaffin cells. Biochem. Pharmacol. 30:855–860.
Schneider, A. S., Cline, H. T., and Lemair, S. 1979. Rapid rise in cyclic AMP accompanies catecholamine secretion in suspensions of isolated adrenal chromaffin cells. Life Sci. 24:1389–1394.
Yanagihara, N., Isosaki, M., Ohuchi, T., and Oka, M. 1979. Muscarinic receptor-mediated increase in cyclic GMP in isolated bevine adrenal medullary cells. FEBS Lett. 105:296–298.
Burgoyne, R. D. 1991. Control of exocytosis in adrenal chromaffin cells. Biochim. Biophys. Acta 1071:174–202.
Fosberg, E. J., Rojas, E., and Pollard, H. B. 1986. Muscarinic receptor enhancement of nicotine-induced catecholamine secretion may be mediated by phosphoinositide metabolism in bovine adrenal cromaffin cells. J. Biol. Chem. 261:4915–4920.
Kilpatrick, D. L., Ledbetter, F. H., Carson, K. A., Kirshner, A. G., Slepetis, R., and Kirshner, N. 1980. Stability of bovine adrenal medulla cells in culture. J. Neurochem. 35:679–692.
Cheek, T. R., and Burgoyne, R. D. 1985. Effect of activation of muscarinic receptors in intracellular free calcium and secretion in bovine adrenal chromaffin cells. Biochim. Biophys. Acta 846:167–173.
Shirvan, M. H., Pollard, H. B. and Heldman, E. 1991. Mixed nicotinic and muscarinic features of cholinergic receptors coupled to secretion in bovine chromaffin cells. Proc. Natl. Acad. Sci. (U.S.A.) 88:4860–4864.
Bonner, T. I. 1989. The molecular basis of muscarinic receptor diversity. Trends Neurosci. 12:148–151.
Barnard, E. A. 1988. Separating receptors subtypes from their shadow. Nature (News and Views) 335:301–302.
Buckley, N. J., Bonner, T. I., Buckley, C. M., and Brann, M. R. 1989. Antagonist binding properties of five cloned muscarinic receptors expressed in CHO-K1 cells Mol. Pharmacol. 35:469–473.
Peralta, E. G., Ashkenazi, A., Winslow, J. W., Smith, D. H., Ramachandran, J., and Capon, D. J. 1987. Distinct primary structures, ligang-binding properties and tissue-specific expression of four human muscarinic acetylcholine receptors. EMBO J. 6:3923–3929.
Lowry, O. M., Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurement by the Folin phenol reagent. J. Biol. Chem. 193:265–275.
Munson, P. J., and Rodbard, D. 1980. LIGAND: a versatile computerized approach for characterization of ligand binding systems. Anal. Biochem. 107:220–239.
Fernando, J. C. R., Abdallah, E.-S., Evinger, M., Forray, C., and El-Pakahani E. E. 1991. The presence of a m4 subtype muscarinic receptor in the bovine adrenal medulla revealed by mRNA and receptor binding analysis. Eur. J. Pharmacol. Mol. Pharmacol. 207:297–303.
Aguilar, J. S., Criado, M., and De Robertis, E. 1979. Pre- and postsynaptic localization of central muscarinic receptors. Eur. J. Pharmacol. 57:227–230.
Fonseca, M. I., Aguilar, J. S., Skorupa, A. F., and Klein, W. L. 1991. Cellular mapping of m2 muscarinic receptor in rat olfactory bulb using an antiserum raised aginst a cytoplasmic loop peptide. Brain Res., in press.
Cheng, Y. C., and Prusoff, W. H. 1973. Relationship between the inhibition constant (Ki) and the concentration of inhibitor which causes 50% inhibition (150) of an exzymatic reaction. Biochem. Pharmacol. 22:3099–3106.
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Aguilar, J.S., Ballesta, J.J., Reig, J.A. et al. Muscarinic receptor subtypes in bovine adrenal medulla. Neurochem Res 17, 1235–1239 (1992). https://doi.org/10.1007/BF00968406
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DOI: https://doi.org/10.1007/BF00968406