Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1977 Oct 1;75(1):43–55. doi: 10.1083/jcb.75.1.43

Isolation of pure cholinergic nerve endings from Torpedo electric organ. Evaluation of their metabolic properties

PMCID: PMC2111552  PMID: 914896

Abstract

Pure cholinergic nerve endings (synaptosomes) were isolated from the electric organ of Torpedo by a rapid procedure. These synaptosomes are approximately 3 micron in diameter. They contain an occasional mitochondrion, numerous synaptic vesicles, and sometimes an active zone is observed. No postynaptic membrane attachment is found. This nerve ending fraction is extremely pure as shown by morphological controls and biochemical data. It is rich in choline acetyltransferase (450 nmol/h per mg protein) and acetylcholine (ACh) (130 nmol/mg protein). The isolated endings retain their cytoplasmic components and they synthesize ACh and are stable in vitro for several hours, as shown by biochemical measurements and morphological analysis.

Full Text

The Full Text of this article is available as a PDF (4.1 MB).

Selected References

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

  1. DE ROBERTIS E., PELLEGRINO DE IRALDI A., RODRIGUEZ G., GOMEZ C. J. On the isolation of nerve endings and synaptic vesicles. J Biophys Biochem Cytol. 1961 Jan;9:229–235. doi: 10.1083/jcb.9.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dowdall M. J., Barrantes F. J., Stender W., Jovin T. M. Inhibitory action of 1-pyrene butyrylcholine and related compounds on choline uptake by cholinergic nerve endings. J Neurochem. 1976 Nov;27(5):1253–1255. doi: 10.1111/j.1471-4159.1976.tb00337.x. [DOI] [PubMed] [Google Scholar]
  3. Dowdall M. J., Simon E. J. Comparative studies on synaptosomes: uptake of (N-Me-3H)choline by synaptosomes from squid optic lobes. J Neurochem. 1973 Oct;21(4):969–982. doi: 10.1111/j.1471-4159.1973.tb07541.x. [DOI] [PubMed] [Google Scholar]
  4. Dunant Y., Gautron J., Israël M., Lesbats B., Manaranche R. Les compartiments d'acetylcholine de l'organe electrique de la torpille et leurs modifications par la stimulation. J Neurochem. 1972 Aug;19(8):1987–2002. doi: 10.1111/j.1471-4159.1972.tb01488.x. [DOI] [PubMed] [Google Scholar]
  5. Dunant Y., Israël M., Lesbats B., Manaranche R. Loss of vesicular acetycholine in the Torpedo electric organ on discharge against high external resistance. J Neurochem. 1976 Oct;27(4):975–977. doi: 10.1111/j.1471-4159.1976.tb05166.x. [DOI] [PubMed] [Google Scholar]
  6. Dunant Y., Israël M., Lesbats B., Manaranche R. Oscillation of acetylcholine during nerve activity in the Torpedo electric organ. Brain Res. 1977 Apr 8;125(1):123–140. doi: 10.1016/0006-8993(77)90364-x. [DOI] [PubMed] [Google Scholar]
  7. ELLMAN G. L., COURTNEY K. D., ANDRES V., Jr, FEATHER-STONE R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961 Jul;7:88–95. doi: 10.1016/0006-2952(61)90145-9. [DOI] [PubMed] [Google Scholar]
  8. FRONTALI N., TOSCHI G. Subcellular fractions from the electric tissue of Torpedo; morphological aspect and cholinesterase content. Exp Cell Res. 1958 Oct;15(2):446–450. doi: 10.1016/0014-4827(58)90055-7. [DOI] [PubMed] [Google Scholar]
  9. Fonnum F. A rapid radiochemical method for the determination of choline acetyltransferase. J Neurochem. 1975 Feb;24(2):407–409. doi: 10.1111/j.1471-4159.1975.tb11895.x. [DOI] [PubMed] [Google Scholar]
  10. Fonnum F. Isolation of choline esters from aqueous solutions by extraction with sodium tetraphenylboron in organic solvents. Biochem J. 1969 Jun;113(2):291–298. doi: 10.1042/bj1130291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guyenet P., Lefresne P., Rossier J., Beaujouan J. C., Glowinski J. Effect of sodium, hemicholinium-3 and antiparkinson drugs on (14C)acetylcholine synthesis and (3H)choline uptake in rat striatal synaptosomes. Brain Res. 1973 Nov 23;62(2):523–529. doi: 10.1016/0006-8993(73)90717-8. [DOI] [PubMed] [Google Scholar]
  12. HEMSWORTH B. A., MORRIS D. A COMPARISON OF THE N-ALKYL GROUP SPECIFICITY OF CHOLINE ACETYLTRANSFERASE FROM DIFFERENT SPECIES. J Neurochem. 1964 Nov;11:793–803. doi: 10.1111/j.1471-4159.1964.tb06728.x. [DOI] [PubMed] [Google Scholar]
  13. Haga T., Noda H. Choline uptake systems of rat brain synaptosomes. Biochim Biophys Acta. 1973 Jan 26;291(2):564–575. doi: 10.1016/0005-2736(73)90508-7. [DOI] [PubMed] [Google Scholar]
  14. Israel M., Gautron J., Lesbats B. Isolement des vésicules synaptiques de l'organe électrique de la Torpille et localisation de l'acétylcholine à leur niveau. C R Acad Sci Hebd Seances Acad Sci D. 1968 Jan 15;266(3):273–275. [PubMed] [Google Scholar]
  15. Israël M., Gautron J., Lesbats B. Fractionnement de l'organe electrique de la torpille: localisation subcellulaire de l'acetylcholine. J Neurochem. 1970 Oct;17(10):1441–1450. doi: 10.1111/j.1471-4159.1970.tb00511.x. [DOI] [PubMed] [Google Scholar]
  16. Israël M. Localisation de l'acétylcholine des synapses myoneurales et nerf-électroplaque. Arch Anat Microsc Morphol Exp. 1970 Apr-Jun;59(2):67–98. [PubMed] [Google Scholar]
  17. Israël M., Manaranche R., Mastour-Frachon P., Morel N. Isolation of pure cholinergic nerve endings from the electric organ of Torpedo marmorata. Biochem J. 1976 Oct 15;160(1):113–115. doi: 10.1042/bj1600113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Israël M., Tucek S. Utilization of acetate and pyruvate for the synthesis of 'total', 'bound' and 'free' acetylcholine in the electric organ of Torpedo. J Neurochem. 1974 Apr;22(4):487–491. doi: 10.1111/j.1471-4159.1974.tb06883.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Marchbanks R. M., Israël M. Aspects of acetylcholine metabolism in the electric organ of Torpedo marmorata. J Neurochem. 1971 Mar;18(3):439–448. doi: 10.1111/j.1471-4159.1971.tb11971.x. [DOI] [PubMed] [Google Scholar]
  22. Marchbanks R. M., Israël M. The heterogeneity of bound acetylcholine and synaptic vesicles. Biochem J. 1972 Oct;129(5):1049–1061. doi: 10.1042/bj1291049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Michaelson D. M., Sokolovsky M. Neurotransmitter release from viable purely cholinergic Torpedo synaptosomes. Biochem Biophys Res Commun. 1976 Nov 8;73(1):25–31. doi: 10.1016/0006-291x(76)90492-7. [DOI] [PubMed] [Google Scholar]
  24. Morel N. Effect of choline on the rates of synthesis and of release of acetylcholine in the electric organ of Torpedo. J Neurochem. 1976 Sep;27(3):779–784. doi: 10.1111/j.1471-4159.1976.tb10408.x. [DOI] [PubMed] [Google Scholar]
  25. SHERIDAN M. N. THE FINE STRUCTURE OF THE ELECTRIC ORGAN OF TORPEDO MARMORATA. J Cell Biol. 1965 Jan;24:129–141. doi: 10.1083/jcb.24.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Whittaker V. P., Essman W. B., Dowe G. H. The isolation of pure cholinergic synaptic vesicles from the electric organs of elasmobranch fish of the family Torpedinidae. Biochem J. 1972 Jul;128(4):833–845. doi: 10.1042/bj1280833. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES