Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1989 Jan;408:233–249. doi: 10.1113/jphysiol.1989.sp017457

The effect of anions on bound acetylcholine in frog sartorius muscle.

B Ceccarelli 1, P C Molenaar 1, B S Oen 1, R L Polak 1, F Torri-Tarelli 1, G T van Kempen 1
PMCID: PMC1190401  PMID: 2789283

Abstract

1. Frog sartorius muscles were treated with an irreversible cholinesterase inhibitor and then incubated in isotonic potassium propionate solution (isotonic KPr). Total and bound, presumably vesicular, acetylcholine (ACh) in the tissue and ACh in the medium were assayed by mass fragmentography, miniature end-plate potentials (MEPPs) were recorded and the end-plates were investigated by electron microscopy. 2. Incubation in isotonic KPr for 30 min stimulated ACh release and concomitantly decreased total and bound ACh. Nerve stimulation for 30 min by trains of impulses (0.1 s trains of 100 Hz, 1 train s-1) in normal-potassium propionate-containing solution had the same effects. 3. When the tissue was incubated in normal-K+ Ringer solution for 3 h, following chemical or electric stimulation, bound ACh recovered to about 75% of the initial value, provided that Cl- ions were present in the medium. In the presence of propionate instead of Cl- ions almost no recovery of bound ACh took place. There was also recovery of bound ACh in the presence of either NO3- or gluconate ions. In NO3- it was the same as in Cl-, but in gluconate it was less than found in Cl- -containing medium. 4. Recovery of total ACh, in contrast to bound ACh, took place even in the presence of propionate ions, showing that extracellular Cl- is not required for the synthesis of ACh. 5. In terminals recovered in normal Ringer solution, many synaptic vesicles were found, but terminals 'recovered' in propionate solution were depleted of vesicles. 6. From these and other results it is concluded that the recycling of synaptic vesicles normally requires the presence of extracellular chloride.

Full text

PDF
233

Images in this article

Selected References

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

  1. Ceccarelli B., Fesce R., Grohovaz F., Haimann C. The effect of potassium on exocytosis of transmitter at the frog neuromuscular junction. J Physiol. 1988 Jul;401:163–183. doi: 10.1113/jphysiol.1988.sp017156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ceccarelli B., Grohovaz F., Hurlbut W. P. Freeze-fracture studies of frog neuromuscular junctions during intense release of neurotransmitter. II. Effects of electrical stimulation and high potassium. J Cell Biol. 1979 Apr;81(1):178–192. doi: 10.1083/jcb.81.1.178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ceccarelli B., Hurlbut W. P., Mauro A. Depletion of vesicles from frog neuromuscular junctions by prolonged tetanic stimulation. J Cell Biol. 1972 Jul;54(1):30–38. doi: 10.1083/jcb.54.1.30. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Couteaux R., Pécot-Dechavassine M. Vésicules synaptiques et poches au niveau des "zones actives" de la jonction neuromusculaire. C R Acad Sci Hebd Seances Acad Sci D. 1970 Dec 21;271(25):2346–2349. [PubMed] [Google Scholar]
  5. Gennaro J. F., Jr, Nastuk W. L., Rutherford D. T. Reversible depletion of synaptic vesicles induced by application of high external potassium to the frog neuromuscular junction. J Physiol. 1978 Jul;280:237–247. doi: 10.1113/jphysiol.1978.sp012382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Heuser J. E. Proceedings: A possible origin of the 'giant' spontaneous potentials that occur after prolonged transmitter release at frog neuromuscular junctions. J Physiol. 1974 Jun;239(2):106P–108P. doi: 10.1113/jphysiol.1974.sp010593. [DOI] [PubMed] [Google Scholar]
  7. Heuser J. E., Reese T. S. Evidence for recycling of synaptic vesicle membrane during transmitter release at the frog neuromuscular junction. J Cell Biol. 1973 May;57(2):315–344. doi: 10.1083/jcb.57.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Johnson R. G., Jr Proton pumps and chemiosmotic coupling as a generalized mechanism for neurotransmitter and hormone transport. Ann N Y Acad Sci. 1987;493:162–177. doi: 10.1111/j.1749-6632.1987.tb27198.x. [DOI] [PubMed] [Google Scholar]
  9. KATZ B., MILEDI R. PROPAGATION OF ELECTRIC ACTIVITY IN MOTOR NERVE TERMINALS. Proc R Soc Lond B Biol Sci. 1965 Feb 16;161:453–482. doi: 10.1098/rspb.1965.0015. [DOI] [PubMed] [Google Scholar]
  10. MILEDI R. The acetylcholine sensitivity of frog muscle fibres after complete or partial devervation. J Physiol. 1960 Apr;151:1–23. [PMC free article] [PubMed] [Google Scholar]
  11. Mallart A., Dreyer F., Peper K. Current-voltage relation and reversal potential at junctional and extrajunctional ACh-receptors of the frog neuromuscular junction. Pflugers Arch. 1976 Mar 11;362(1):43–47. doi: 10.1007/BF00588679. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Miledi R., Molenaar P. C., Polak R. L. An analysis of acetylcholine in frog muscle by mass fragmentography. Proc R Soc Lond B Biol Sci. 1977 Jun 15;197(1128):285–297. doi: 10.1098/rspb.1977.0071. [DOI] [PubMed] [Google Scholar]
  14. Miledi R., Molenaar P. C., Polak R. L. Free and bound acetylcholine in frog muscle. J Physiol. 1982 Dec;333:189–199. doi: 10.1113/jphysiol.1982.sp014448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Miledi R., Molenaar P. C., Polak R. L. The effect of lanthanum ions on acetylcholine in frog muscle. J Physiol. 1980 Dec;309:199–214. doi: 10.1113/jphysiol.1980.sp013504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Molenaar P. C., Oen B. S., Polak R. L. Effect of chloride ions on giant miniature end-plate potentials at the frog neuromuscular junction. J Physiol. 1987 Feb;383:143–152. doi: 10.1113/jphysiol.1987.sp016401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Molenaar P. C., Polak R. L. Potassium propionate causes preferential loss of 'bound' acetylcholine in frog muscle. Neurosci Lett. 1983 Dec 30;43(2-3):209–213. doi: 10.1016/0304-3940(83)90189-1. [DOI] [PubMed] [Google Scholar]
  18. Polak R. L., Molenaar P. C. A method for determination of acetylcholine by slow pyrolysis combined with mass fragmentography on a packed capillary column. J Neurochem. 1979 Feb;32(2):407–412. doi: 10.1111/j.1471-4159.1979.tb00364.x. [DOI] [PubMed] [Google Scholar]
  19. Polak R. L., Molenaar P. C. Pitfalls in determination of acetylcholine from brain by pyrolysis-gas chromatography/mass spectrometry. J Neurochem. 1974 Dec;23(6):1295–1297. doi: 10.1111/j.1471-4159.1974.tb12230.x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES