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. 1974 May;71(5):1725–1729. doi: 10.1073/pnas.71.5.1725

Muscarinic Cholinergic Binding in Rat Brain

Henry I Yamamura 1,2,3,4, Solomon H Snyder 1,2,3,4,†
PMCID: PMC388311  PMID: 4151898

Abstract

Binding sites with high affinity and specificity for [3H]quinuclidinyl benzilate (QNB) are present in homogenates of rat brain. The characteristics of the binding sites resemble those of muscarinic cholinergic receptors. Specific binding is saturable with respect to [3H]QNB and tissue concentration and is time-, temperature-, and pH-dependent. The bimolecular rate of association (2.0 × 108 M-1 min-1) and dissociation (1.2 × 10-2 min-1) at 35° indicate a dissociation constant of 60 pM and a density of 65 pmol/g of brain. Muscarinic antagonists and agonists displace specific [3H]QNB binding, while nicotinic and non-cholinergic drugs possess little affinity for [3H]QNB-binding sites.

Keywords: quinuclidinyl benzilate, receptors

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albanus L. Central and peripheral effects of anticholinergic compounds. Acta Pharmacol Toxicol (Copenh) 1970;28(4):305–326. doi: 10.1111/j.1600-0773.1970.tb00557.x. [DOI] [PubMed] [Google Scholar]
  2. Berg D. K., Kelly R. B., Sargent P. B., Williamson P., Hall Z. W. Binding of -bungarotoxin to acetylcholine receptors in mammalian muscle (snake venom-denervated muscle-neonatal muscle-rat diaphragm-SDS-polyacrylamide gel electrophoresis). Proc Natl Acad Sci U S A. 1972 Jan;69(1):147–151. doi: 10.1073/pnas.69.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bosmann H. B. Acetylcholine receptor. I. Identification and biochemical characteristics of a cholinergic receptor of guinea pig cerebral cortex. J Biol Chem. 1972 Jan 10;247(1):130–145. [PubMed] [Google Scholar]
  4. Campbell L. B., Jenden D. J. Gas chromatographic evaluation of the influence of oxotremorine upon the regional distribution of acetylcholine in rat brain. J Neurochem. 1970 Dec;17(12):1697–1699. doi: 10.1111/j.1471-4159.1970.tb11396.x. [DOI] [PubMed] [Google Scholar]
  5. Changeux J. P., Kasai M., Lee C. Y. Use of a snake venom toxin to characterize the cholinergic receptor protein. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1241–1247. doi: 10.1073/pnas.67.3.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Robertis E., Fiszer S., Pasquini J. M., Soto E. F. Isolation and chemical nature of the receptor for d-tubocurarine in nerve-ending membranes of the cerebral cortex. J Neurobiol. 1969;1(1):41–52. doi: 10.1002/neu.480010105. [DOI] [PubMed] [Google Scholar]
  7. Eldefrawi A. T., O'Brien R. D. Binding of muscarone by extracts of housefly brain: relationship to receptors for acetylcholine. J Neurochem. 1970 Aug;17(8):1287–1293. doi: 10.1111/j.1471-4159.1970.tb03377.x. [DOI] [PubMed] [Google Scholar]
  8. Eldefrawi M. E., Britten A. G., Eldefrawi A. T. Acetylcholine binding to Torpedo electroplax: relationship to acetylcholine receptors. Science. 1971 Jul 23;173(3994):338–340. doi: 10.1126/science.173.3994.338. [DOI] [PubMed] [Google Scholar]
  9. Fambrough D. M., Hartzell H. C. Acetylcholine receptors: number and distribution at neuromuscular junctions in rat diaphragm. Science. 1972 Apr 14;176(4031):189–191. doi: 10.1126/science.176.4031.189. [DOI] [PubMed] [Google Scholar]
  10. Farrow J. T., O'Brien R. D. Binding of atropine and muscarone to rat brain fractions and its relation to the acetylcholine receptor. Mol Pharmacol. 1973 Jan;9(1):33–40. [PubMed] [Google Scholar]
  11. Fewtrell C., Rang H. P. Distribution of bound 3 H-benzilylcholine mustard in subcellular fractions of smooth muscle from guinea-pig ileum. Br J Pharmacol. 1971 Oct;43(2):417P–418P. [PMC free article] [PubMed] [Google Scholar]
  12. Gill E. W., Rang H. P. An alkylating derivative of benzilylcholine with specific and long-lasting parasympatholytic activity. Mol Pharmacol. 1966 Jul;2(4):284–297. [PubMed] [Google Scholar]
  13. Goldberg A. M., Welch B. L. Adaptation of the adrenal medulla: sustained increase in choline acetyltransferase by psychosocial stimulation. Science. 1972 Oct 20;178(4058):319–320. doi: 10.1126/science.178.4058.319. [DOI] [PubMed] [Google Scholar]
  14. Hartzell H. C., Fambrough D. M. Acetylcholine receptors. Distribution and extrajunctional density in rat diaphragm after denervation correlated with acetylcholine sensitivity. J Gen Physiol. 1972 Sep;60(3):248–262. doi: 10.1085/jgp.60.3.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hiley C. R., Young J. M., Burgen A. S. Labelling of cholinergic receptors in subcellular fractions from rat cerebral cortex. Biochem J. 1972 May;127(5):86P–86P. doi: 10.1042/bj1270086pa. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kebabian J. W., Petzold G. L., Greengard P. Dopamine-sensitive adenylate cyclase in caudate nucleus of rat brain, and its similarity to the "dopamine receptor". Proc Natl Acad Sci U S A. 1972 Aug;69(8):2145–2149. doi: 10.1073/pnas.69.8.2145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klett R. P., Fulpius B. W., Cooper D., Smith M., Reich E., Possani L. D. The acetylcholine receptor. I. Purification and characterization of a macromolecule isolated from Electrophorus electricus. J Biol Chem. 1973 Oct 10;248(19):6841–6853. [PubMed] [Google Scholar]
  18. Krnjević K., Phillis J. W. Pharmacological properties of acetylcholine-sensitive cells in the cerebral cortex. J Physiol. 1963 May;166(2):328–350. doi: 10.1113/jphysiol.1963.sp007107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. MCCAMAN R. E., HUNT J. M. MICRODETERMINATION OF CHOLINE ACETYLASE IN NERVOUS TISSUE. J Neurochem. 1965 Apr;12:253–259. doi: 10.1111/j.1471-4159.1965.tb06762.x. [DOI] [PubMed] [Google Scholar]
  21. McCaman M. W., Tomey L. R., McCaman R. E. Radiomimetric assay of acetylcholinesterase activity in submicrogram amounts of tissue. Life Sci. 1968 Mar 15;7(6):233–244. doi: 10.1016/0024-3205(68)90017-9. [DOI] [PubMed] [Google Scholar]
  22. Meyerhöffer A. Absolute configuration of 3-quinuclidinyl benzilate and the behavioral effect in the dog of the optical isomers. J Med Chem. 1972 Sep;15(9):994–995. doi: 10.1021/jm00279a030. [DOI] [PubMed] [Google Scholar]
  23. Miledi R., Molinoff P., Potter L. T. Isolation of the cholinergic receptor protein of Torpedo electric tissue. Nature. 1971 Feb 19;229(5286):554–557. doi: 10.1038/229554a0. [DOI] [PubMed] [Google Scholar]
  24. Miledi R., Potter L. T. Acetylcholine receptors in muscle fibres. Nature. 1971 Oct 29;233(5322):599–603. doi: 10.1038/233599a0. [DOI] [PubMed] [Google Scholar]
  25. O'Brien R. D., Eldefrawi M. E., Eldefrawi A. T. Isolation of acetylcholine receptors. Annu Rev Pharmacol. 1972;12:19–34. doi: 10.1146/annurev.pa.12.040172.000315. [DOI] [PubMed] [Google Scholar]
  26. O'Brien R. D., Gilmour L. P. A muscarone-binding material in electroplax and its relation to the acetylcholine receptor. I. Centrifugal assay. Proc Natl Acad Sci U S A. 1969 Jun;63(2):496–503. doi: 10.1073/pnas.63.2.496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. PATON W. D., RANG H. P. THE UPTAKE OF ATROPINE AND RELATED DRUGS BY INTESTINAL SMOOTH MUSCLE OF THE GUINEA-PIG IN RELATION TO ACETYLCHOLINE RECEPTORS. Proc R Soc Lond B Biol Sci. 1965 Aug 24;163:1–44. doi: 10.1098/rspb.1965.0058. [DOI] [PubMed] [Google Scholar]
  28. Pert C. B., Snyder S. H. Opiate receptor: demonstration in nervous tissue. Science. 1973 Mar 9;179(4077):1011–1014. doi: 10.1126/science.179.4077.1011. [DOI] [PubMed] [Google Scholar]
  29. Pert C. B., Snyder S. H. Properties of opiate-receptor binding in rat brain. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2243–2247. doi: 10.1073/pnas.70.8.2243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Raftery M. A., Schmidt J., Clark D. G., Wolcott R. G. Demonstration of a specific -bungarotoxin binding component in electrophorus electricus electroplax membranes. Biochem Biophys Res Commun. 1971 Dec 17;45(6):1622–1629. doi: 10.1016/0006-291x(71)90207-5. [DOI] [PubMed] [Google Scholar]
  31. Schmidt D. E., Speth R. C., Welsch F., Schmidt M. J. The use of microwave radiation in the determination of acetylcholine in the rat brain. Brain Res. 1972 Mar 24;38(2):377–389. doi: 10.1016/0006-8993(72)90720-2. [DOI] [PubMed] [Google Scholar]
  32. Sethy V. H., Roth R. H., Kuhar M. J., Van Woert M. H. Choline and acetylcholine: regional distribution and effect of degeneration of cholinergic nerve terminals in the rat hippocampus. Neuropharmacology. 1973 Sep;12(9):819–823. doi: 10.1016/0028-3908(73)90034-8. [DOI] [PubMed] [Google Scholar]
  33. Yamamura H. I., Snyder S. H. Choline: high-affinity uptake by rat brain synaptosomes. Science. 1972 Nov 10;178(4061):626–628. doi: 10.1126/science.178.4061.626. [DOI] [PubMed] [Google Scholar]
  34. Yamamura H. I., Snyder S. H. High affinity transport of choline into synaptosomes of rat brain. J Neurochem. 1973 Dec;21(6):1355–1374. doi: 10.1111/j.1471-4159.1973.tb06022.x. [DOI] [PubMed] [Google Scholar]
  35. Young A. B., Snyder S. H. Strychnine binding associated with glycine receptors of the central nervous system. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2832–2836. doi: 10.1073/pnas.70.10.2832. [DOI] [PMC free article] [PubMed] [Google Scholar]

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