Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1969 Jun;113(1):97–107. doi: 10.1042/bj1130097

The subcellular distribution of [N-Me-3H]-acetylcholine synthesized by brain in vivo

L W Chakrin 1,*, V P Whittaker 1
PMCID: PMC1184608  PMID: 5806399

Abstract

1. Three forms of acetylcholine occur in subcellular fractions of brain tissue: free acetylcholine, present in the high-speed supernatant from eserinized sucrose homogenates; stable bound acetylcholine, present in synaptic vesicles; and labile bound acetylcholine, present in the cytoplasm of synaptosomes (detached presynaptic nerve terminals). 2. The relationship between these forms has been investigated by isolating the subcellular fractions from the cortical tissue of cats and guinea pigs excised 1hr. after infiltration of [N-Me-3H]choline into the cortex in vivo. 3. Since choline is a ubiquitous metabolite, means were devised for isolating the radioactive acetylcholine on columns of the weak acid ion-exchange resin IRF-97; control experiments with samples of extracts treated with acetylcholinesterase showed that the radioactivity attributed to acetylcholine migrated to the choline peak after cholinesterase treatment. 4. The specific radioactivities of the various forms of acetylcholine were different: labile bound (synaptosomal cytoplasmic) acetylcholine had the highest, stable bound (vesicular) acetylcholine the next highest, and the high-speed-supernatant form the lowest. 5. It is concluded that the various forms of acetylcholine could not have arisen during fractionation from a single pre-existing pool of acetylcholine.

Full text

PDF
97

Selected References

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

  1. APPLETON H. D., LA DU B. N., Jr, LEVY B., STEELE J. M., BRODIE B. B. A chemical method for the determination of free choline in plasma. J Biol Chem. 1953 Dec;205(2):803–813. [PubMed] [Google Scholar]
  2. Chakrin L. W., Shideman F. E., Marrazzi A. S. The in vivo synthesis and release of tritium labeled acetylcholine by cat cerebral cortex. Int J Neuropharmacol. 1968 Jul;7(4):351–358. doi: 10.1016/0028-3908(68)90017-8. [DOI] [PubMed] [Google Scholar]
  3. Chakrin L. W., Shideman F. E. Synthesis of acetylcholine from labeled choline by brain. Int J Neuropharmacol. 1968 Jul;7(4):337–349. doi: 10.1016/0028-3908(68)90016-6. [DOI] [PubMed] [Google Scholar]
  4. DOMER F. R., FELDBERG W. Scratching movements and facilitation of the scratch reflex produced by tubocurarine in cats. J Physiol. 1960 Aug;153:35–51. doi: 10.1113/jphysiol.1960.sp006517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dudley H. W. Observations on acetylcholine. Biochem J. 1929;23(5):1064–1074. doi: 10.1042/bj0231064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fonnum F. Choline acetyltransferase binding to and release from membranes. Biochem J. 1968 Sep;109(3):389–398. doi: 10.1042/bj1090389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fonnum F. The 'compartmentation' of choline acetyltransferase within the synaptosome. Biochem J. 1967 Apr;103(1):262–270. doi: 10.1042/bj1030262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. GARDINER J. E., WITTAKER V. P. The identification of propionycholine as a constituent of ox spleen. Biochem J. 1954 Sep;58(1):24–29. doi: 10.1042/bj0580024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HEBB C. O., WHITTAKER V. P. Intracellular distributions of acetylcholine and choline acetylase. J Physiol. 1958 Jun 18;142(1):187–196. doi: 10.1113/jphysiol.1958.sp006008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. JOHNSON M. K., WHITTAKER V. P. LACTATE DEHYDROGENASE AS A CYTOPLASMIC MARKER IN BRAIN. Biochem J. 1963 Sep;88:404–409. doi: 10.1042/bj0880404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. LUFT J. H. Permanganate; a new fixative for electron microscopy. J Biophys Biochem Cytol. 1956 Nov 25;2(6):799–802. doi: 10.1083/jcb.2.6.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lewis P. R., Shute C. C. The distribution of cholinesterase in cholinergic neurons demonstrated with the electron microscope. J Cell Sci. 1966 Sep;1(3):381–390. doi: 10.1242/jcs.1.3.381. [DOI] [PubMed] [Google Scholar]
  14. Marchbanks R. M. Exchangeability of radioactive acetylcholine with the bound acetylcholine of synaptosomes and synaptic vesicles. Biochem J. 1968 Jan;106(1):87–95. doi: 10.1042/bj1060087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marchbanks R. M. The osmotically sensitive potassium and sodium compartments of synaptosomes. Biochem J. 1967 Jul;104(1):148–157. doi: 10.1042/bj1040148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McCaman R. E., Rodríguez de Lores G., De Robertis E. Species differences in subcellular distribution of choline acetylase in the CNS. A study of choline acetylase, acetylcholinesterase, 5-hydroxytryptophan decarboxylase, and monoamine oxidase in four species. J Neurochem. 1965 Nov;12(11):927–935. doi: 10.1111/j.1471-4159.1965.tb11936.x. [DOI] [PubMed] [Google Scholar]
  17. Silver A. Cholinesterases of the central nervous system with special reference to the cerebellum. Int Rev Neurobiol. 1967;10:57–109. doi: 10.1016/s0074-7742(08)60151-8. [DOI] [PubMed] [Google Scholar]
  18. Tucek S. Subcellular distribution of acetyl-CoA synthetase, ATP citrate lyase, citrate synthase, choline acetyltransferase, fumarate hydratase and lactate dehydrogenase in mammalian brain tissue. J Neurochem. 1967 May;14(5):531–545. doi: 10.1111/j.1471-4159.1967.tb09552.x. [DOI] [PubMed] [Google Scholar]
  19. WHITTAKER V. P., SHERIDAN M. N. THE MORPHOLOGY AND ACETYLCHOLINE CONTENT OF ISOLATED CEREBRAL CORTICAL SYNAPTIC VESICLES. J Neurochem. 1965 May;12:363–372. doi: 10.1111/j.1471-4159.1965.tb04237.x. [DOI] [PubMed] [Google Scholar]
  20. WHITTAKER V. P. The isolation and characterization of acetylcholine-containing particles from brain. Biochem J. 1959 Aug;72:694–706. doi: 10.1042/bj0720694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wallach M. B., Goldberg A. M., Shideman F. E. The synthesis of labeled acetylcholine by the isolated cat heart and its release by vagal stimulation. Int J Neuropharmacol. 1967 Jul;6(4):317–323. doi: 10.1016/0028-3908(67)90021-4. [DOI] [PubMed] [Google Scholar]
  22. Whittaker V. P., Michaelson I. A., Kirkland R. J. The separation of synaptic vesicles from nerve-ending particles ('synaptosomes'). Biochem J. 1964 Feb;90(2):293–303. doi: 10.1042/bj0900293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Whittaker V. P. The storage of transmitters in the central nervous system. Biochem J. 1968 Sep;109(3):20P–21P. doi: 10.1042/bj1090020p. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES