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. 1973 May 1;57(2):499–524. doi: 10.1083/jcb.57.2.499

TURNOVER OF TRANSMITTER AND SYNAPTIC VESICLES AT THE FROG NEUROMUSCULAR JUNCTION

B Ceccarelli 1, W P Hurlbut 1, A Mauro 1
PMCID: PMC2108980  PMID: 4348791

Abstract

Curarized cutaneous pectoris nerve-muscle preparations from frogs were stimulated at 10/s or at 2/s for periods ranging from 20 min to 4 h. End plate potential were recorded intracellularly and used to estimate the quantity of transmitter secreted during the period of stimulation. At the ends of the periods of stimulation the preparations were either fixed for electron microscopy or treated with black widow spider venom to determine the quantities of transmitter remainind in the terminal. Horseradish peroxidase or dextran was added to the bathing solution and used as a tracer to detect the formation of vesicles from the axolemma. During 4 h of stimulation at 2/s many new vesicles were formed from the axolemma and the quantity of transmitter secreted was several times greater than the quantity in the initial store. After this period of stimulation, the terminals were severely depleted of transmitter, but not of vesicles, and their general morphological organization was normal. During 20 min of stimulation at 10/s the nerve terminals swelled and were severely depleted both of vesicles and of transmitter. During a subsequent hour of rest the changes in morphology were largely reversed, many new vesicles were formed from the axolemma and the stores of transmitter were partially replenished. These results suggest (a) that synaptic vesicles fuse with, and re-form from, the membrane of the nerve terminal during and after stimulation and (b), that the re-formed vesicles can store and release transmitter.

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Selected References

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

  1. Atwood H. L., Lang F., Morin W. A. Synaptic vesicles: selective depletion in crayfish excitatory and inhibitory axons. Science. 1972 Jun 23;176(4041):1353–1355. doi: 10.1126/science.176.4041.1353. [DOI] [PubMed] [Google Scholar]
  2. Auerbach A., Betz W. Does curare affect transmitter release? J Physiol. 1971 Mar;213(3):691–705. doi: 10.1113/jphysiol.1971.sp009409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BIRKS R., HUXLEY H. E., KATZ B. The fine structure of the neuromuscular junction of the frog. J Physiol. 1960 Jan;150:134–144. doi: 10.1113/jphysiol.1960.sp006378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bittner G. D., Kennedy D. Quantitative aspects of transmitter release. J Cell Biol. 1970 Dec;47(3):585–592. doi: 10.1083/jcb.47.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Capek R., Esplin D. W., Salehmoghaddam S. Rates of transmitter turnover at the frog neuromuscular junction estimated by electrophysiological techniques. J Neurophysiol. 1971 Sep;34(5):831–841. doi: 10.1152/jn.1971.34.5.831. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Clark A. W., Hurlbut W. P., Mauro A. Changes in the fine structure of the neuromuscular junction of the frog caused by black widow spider venom. J Cell Biol. 1972 Jan;52(1):1–14. doi: 10.1083/jcb.52.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. DEL CASTILLO J., KATZ B. Quantal components of the end-plate potential. J Physiol. 1954 Jun 28;124(3):560–573. doi: 10.1113/jphysiol.1954.sp005129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  11. Holtzman E. Cytochemical studies of protein transport in the nervous system. Philos Trans R Soc Lond B Biol Sci. 1971 Jun 17;261(839):407–421. doi: 10.1098/rstb.1971.0075. [DOI] [PubMed] [Google Scholar]
  12. Holtzman E., Freeman A. R., Kashner L. A. Stimulation-dependent alterations in peroxidase uptake at lobster neuromuscular junctions. Science. 1971 Aug 20;173(3998):733–736. doi: 10.1126/science.173.3998.733. [DOI] [PubMed] [Google Scholar]
  13. Hubbard J. I., Kwanbunbumpen S. Evidence for the vesicle hypothesis. J Physiol. 1968 Feb;194(2):407–420. doi: 10.1113/jphysiol.1968.sp008415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hubbard J. I., Wilson D. F., Miyamoto M. Reduction of transmitter release by D-tubocurarine. Nature. 1969 Aug 2;223(5205):531–533. doi: 10.1038/223531a0. [DOI] [PubMed] [Google Scholar]
  15. JENKINSON D. H. The antagonism between tubocurarine and substances which depolarize the motor end-plate. J Physiol. 1960 Jul;152:309–324. doi: 10.1113/jphysiol.1960.sp006489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Longenecker H. E., Jr, Hurlbut W. P., Mauro A., Clark A. W. Effects of black widow spider venom on the frog neuromuscular junction. Effects on end-plate potential, miniature end-plate potential and nerve terminal spike. Nature. 1970 Feb 21;225(5234):701–703. doi: 10.1038/225701a0. [DOI] [PubMed] [Google Scholar]
  17. OTSUKA M., ENDO M., NONOMURA Y. Presynaptic nature of neuromuscular depression. Jpn J Physiol. 1962 Dec 15;12:573–584. doi: 10.2170/jjphysiol.12.573. [DOI] [PubMed] [Google Scholar]
  18. Perri V., Sacchi O., Raviola E., Raviola G. Evaluation of the number and distribution of synaptic vesicles at cholinergic nerve-endings after sustained stimulation. Brain Res. 1972 Apr 28;39(2):526–529. doi: 10.1016/0006-8993(72)90458-1. [DOI] [PubMed] [Google Scholar]
  19. Pysh J. J., Wiley R. G. Morphologic alterations of synapses in electrically stimulated superior cervical ganglia of the cat. Science. 1972 Apr 14;176(4031):191–193. doi: 10.1126/science.176.4031.191. [DOI] [PubMed] [Google Scholar]
  20. Simionescu N., Palade G. E. Dextrans and glycogens as particulate tracers for studying capillary permeability. J Cell Biol. 1971 Sep;50(3):616–624. doi: 10.1083/jcb.50.3.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Simionescu N., Simionescu M., Palade G. E. Permeability of intestinal capillaries. Pathway followed by dextrans and glycogens. J Cell Biol. 1972 May;53(2):365–392. doi: 10.1083/jcb.53.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. TAKEUCHI A., TAKEUCHI N. Active phase of frog's end-plate potential. J Neurophysiol. 1959 Jul;22(4):395–411. doi: 10.1152/jn.1959.22.4.395. [DOI] [PubMed] [Google Scholar]

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