Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Oct;85(19):7366–7370. doi: 10.1073/pnas.85.19.7366

Differential effect of alpha-latrotoxin on exocytosis from small synaptic vesicles and from large dense-core vesicles containing calcitonin gene-related peptide at the frog neuromuscular junction.

M Matteoli 1, C Haimann 1, F Torri-Tarelli 1, J M Polak 1, B Ceccarelli 1, P De Camilli 1
PMCID: PMC282187  PMID: 3050995

Abstract

The regulatory peptide called calcitonin gene-related peptide (CGRP) was detected by immunofluorescence in frog motor neurons and motor nerve terminals. In motor nerve terminals, CGRP-like immunoreactivity was found to be segregated within large dense-core vesicles. To determine whether exocytosis from acetylcholine-containing small synaptic vesicles and from CGRP-containing large dense-core vesicles can be independently stimulated, nerve-muscle preparations were exposed to alpha-latrotoxin. This toxin induced complete depletion of acetylcholine-containing small synaptic vesicles but did not induce a parallel depletion of CGRP-like immunoreactivity and of large dense-core vesicles. These effects were independent of the presence of extracellular Ca2+ and occurred both at room temperature and at low temperature (1-3 degrees C). These findings suggest that exocytosis from the two vesicle populations is mediated by distinct biochemical mechanisms, which might be differentially regulated by physiological stimuli.

Full text

PDF
7366

Images in this article

7367Image
on p.7367
7368Image
on p.7368
7368Image
on p.7368
7368Image
on p.7368
7368Image
on p.7368
7368Image
on p.7368
7368Image
on p.7368
7369Image
on p.7369

Selected References

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

  1. Amara S. G., Jonas V., Rosenfeld M. G., Ong E. S., Evans R. M. Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products. Nature. 1982 Jul 15;298(5871):240–244. doi: 10.1038/298240a0. [DOI] [PubMed] [Google Scholar]
  2. Andersson P. O., Bloom S. R., Edwards A. V., Järhult J. Effects of stimulation of the chorda tympani in bursts on submaxillary responses in the cat. J Physiol. 1982 Jan;322:469–483. doi: 10.1113/jphysiol.1982.sp014050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bähler M., Greengard P. Synapsin I bundles F-actin in a phosphorylation-dependent manner. Nature. 1987 Apr 16;326(6114):704–707. doi: 10.1038/326704a0. [DOI] [PubMed] [Google Scholar]
  4. Ceccarelli B., Hurlbut W. P. Ca2+-dependent recycling of synaptic vesicles at the frog neuromuscular junction. J Cell Biol. 1980 Oct;87(1):297–303. doi: 10.1083/jcb.87.1.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ceccarelli B., Hurlbut W. P., Iezzi N. Effect of alpha-latrotoxin on the frog neuromuscular junction at low temperature. J Physiol. 1988 Aug;402:195–217. doi: 10.1113/jphysiol.1988.sp017200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Camilli P., Cameron R., Greengard P. Synapsin I (protein I), a nerve terminal-specific phosphoprotein. I. Its general distribution in synapses of the central and peripheral nervous system demonstrated by immunofluorescence in frozen and plastic sections. J Cell Biol. 1983 May;96(5):1337–1354. doi: 10.1083/jcb.96.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Camilli P., Greengard P. Synapsin I: a synaptic vesicle-associated neuronal phosphoprotein. Biochem Pharmacol. 1986 Dec 15;35(24):4349–4357. doi: 10.1016/0006-2952(86)90747-1. [DOI] [PubMed] [Google Scholar]
  8. Edström A., Hanson M. Temperature effects on fast axonal transport of proteins in vitro in frog sciatic nerves. Brain Res. 1973 Aug 30;58(2):345–354. doi: 10.1016/0006-8993(73)90006-1. [DOI] [PubMed] [Google Scholar]
  9. Fesce R., Segal J. R., Ceccarelli B., Hurlbut W. P. Effects of black widow spider venom and Ca2+ on quantal secretion at the frog neuromuscular junction. J Gen Physiol. 1986 Jul;88(1):59–81. doi: 10.1085/jgp.88.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fontaine B., Klarsfeld A., Changeux J. P. Calcitonin gene-related peptide and muscle activity regulate acetylcholine receptor alpha-subunit mRNA levels by distinct intracellular pathways. J Cell Biol. 1987 Sep;105(3):1337–1342. doi: 10.1083/jcb.105.3.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fontaine B., Klarsfeld A., Hökfelt T., Changeux J. P. Calcitonin gene-related peptide, a peptide present in spinal cord motoneurons, increases the number of acetylcholine receptors in primary cultures of chick embryo myotubes. Neurosci Lett. 1986 Oct 30;71(1):59–65. doi: 10.1016/0304-3940(86)90257-0. [DOI] [PubMed] [Google Scholar]
  12. Frontali N., Ceccarelli B., Gorio A., Mauro A., Siekevitz P., Tzeng M. C., Hurlbut W. P. Purification from black widow spider venom of a protein factor causing the depletion of synaptic vesicles at neuromuscular junctions. J Cell Biol. 1976 Mar;68(3):462–479. doi: 10.1083/jcb.68.3.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibson S. J., Polak J. M., Bloom S. R., Sabate I. M., Mulderry P. M., Ghatei M. A., McGregor G. P., Morrison J. F., Kelly J. S., Evans R. M. Calcitonin gene-related peptide immunoreactivity in the spinal cord of man and of eight other species. J Neurosci. 1984 Dec;4(12):3101–3111. doi: 10.1523/JNEUROSCI.04-12-03101.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Haimann C., Torri-Tarelli F., Fesce R., Ceccarelli B. Measurement of quantal secretion induced by ouabain and its correlation with depletion of synaptic vesicles. J Cell Biol. 1985 Nov;101(5 Pt 1):1953–1965. doi: 10.1083/jcb.101.5.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hirning L. D., Fox A. P., McCleskey E. W., Olivera B. M., Thayer S. A., Miller R. J., Tsien R. W. Dominant role of N-type Ca2+ channels in evoked release of norepinephrine from sympathetic neurons. Science. 1988 Jan 1;239(4835):57–61. doi: 10.1126/science.2447647. [DOI] [PubMed] [Google Scholar]
  16. Hökfelt T., Millhorn D., Seroogy K., Tsuruo Y., Ceccatelli S., Lindh B., Meister B., Melander T., Schalling M., Bartfai T. Coexistence of peptides with classical neurotransmitters. Experientia. 1987 Jul 15;43(7):768–780. doi: 10.1007/BF01945354. [DOI] [PubMed] [Google Scholar]
  17. Ip N. Y., Zigmond R. E. Pattern of presynaptic nerve activity can determine the type of neurotransmitter regulating a postsynaptic event. Nature. 1984 Oct 4;311(5985):472–474. doi: 10.1038/311472a0. [DOI] [PubMed] [Google Scholar]
  18. Keller G. A., Tokuyasu K. T., Dutton A. H., Singer S. J. An improved procedure for immunoelectron microscopy: ultrathin plastic embedding of immunolabeled ultrathin frozen sections. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5744–5747. doi: 10.1073/pnas.81.18.5744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Laufer R., Changeux J. P. Calcitonin gene-related peptide elevates cyclic AMP levels in chick skeletal muscle: possible neurotrophic role for a coexisting neuronal messenger. EMBO J. 1987 Apr;6(4):901–906. doi: 10.1002/j.1460-2075.1987.tb04836.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lynch K. Stimulation-induced reduction of large dense core vesicle numbers in cholinergic motor nerve endings. Brain Res. 1980 Jul 21;194(1):249–254. doi: 10.1016/0006-8993(80)91338-4. [DOI] [PubMed] [Google Scholar]
  21. Mason R. T., Peterfreund R. A., Sawchenko P. E., Corrigan A. Z., Rivier J. E., Vale W. W. Release of the predicted calcitonin gene-related peptide from cultured rat trigeminal ganglion cells. Nature. 1984 Apr 12;308(5960):653–655. doi: 10.1038/308653a0. [DOI] [PubMed] [Google Scholar]
  22. Misler S., Falke L. C. Dependence on multivalent cations of quantal release of transmitter induced by black widow spider venom. Am J Physiol. 1987 Sep;253(3 Pt 1):C469–C476. doi: 10.1152/ajpcell.1987.253.3.C469. [DOI] [PubMed] [Google Scholar]
  23. Mulderry P. K., Ghatei M. A., Rodrigo J., Allen J. M., Rosenfeld M. G., Polak J. M., Bloom S. R. Calcitonin gene-related peptide in cardiovascular tissues of the rat. Neuroscience. 1985 Mar;14(3):947–954. doi: 10.1016/0306-4522(85)90156-3. [DOI] [PubMed] [Google Scholar]
  24. Navone F., Greengard P., De Camilli P. Synapsin I in nerve terminals: selective association with small synaptic vesicles. Science. 1984 Dec 7;226(4679):1209–1211. doi: 10.1126/science.6438799. [DOI] [PubMed] [Google Scholar]
  25. Navone F., Jahn R., Di Gioia G., Stukenbrok H., Greengard P., De Camilli P. Protein p38: an integral membrane protein specific for small vesicles of neurons and neuroendocrine cells. J Cell Biol. 1986 Dec;103(6 Pt 1):2511–2527. doi: 10.1083/jcb.103.6.2511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. New H. V., Mudge A. W. Calcitonin gene-related peptide regulates muscle acetylcholine receptor synthesis. 1986 Oct 30-Nov 5Nature. 323(6091):809–811. doi: 10.1038/323809a0. [DOI] [PubMed] [Google Scholar]
  27. Perney T. M., Hirning L. D., Leeman S. E., Miller R. J. Multiple calcium channels mediate neurotransmitter release from peripheral neurons. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6656–6659. doi: 10.1073/pnas.83.17.6656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rane S. G., Holz G. G., 4th, Dunlap K. Dihydropyridine inhibition of neuronal calcium current and substance P release. Pflugers Arch. 1987 Aug;409(4-5):361–366. doi: 10.1007/BF00583789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rodrigo J., Polak J. M., Terenghi G., Cervantes C., Ghatei M. A., Mulderry P. K., Bloom S. R. Calcitonin gene-related peptide (CGRP)-immunoreactive sensory and motor nerves of the mammalian palate. Histochemistry. 1985;82(1):67–74. doi: 10.1007/BF00502092. [DOI] [PubMed] [Google Scholar]
  30. Rosenfeld M. G., Mermod J. J., Amara S. G., Swanson L. W., Sawchenko P. E., Rivier J., Vale W. W., Evans R. M. Production of a novel neuropeptide encoded by the calcitonin gene via tissue-specific RNA processing. Nature. 1983 Jul 14;304(5922):129–135. doi: 10.1038/304129a0. [DOI] [PubMed] [Google Scholar]
  31. Segal J. R., Ceccarelli B., Fesce R., Hurlbut W. P. Miniature endplate potential frequency and amplitude determined by an extension of Campbell's theorem. Biophys J. 1985 Feb;47(2 Pt 1):183–202. doi: 10.1016/s0006-3495(85)83891-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Takami K., Kawai Y., Uchida S., Tohyama M., Shiotani Y., Yoshida H., Emson P. C., Girgis S., Hillyard C. J., MacIntyre I. Effect of calcitonin gene-related peptide on contraction of striated muscle in the mouse. Neurosci Lett. 1985 Sep 30;60(2):227–230. doi: 10.1016/0304-3940(85)90248-4. [DOI] [PubMed] [Google Scholar]
  33. Valtorta F., Villa A., Jahn R., De Camilli P., Greengard P., Ceccarelli B. Localization of synapsin I at the frog neuromuscular junction. Neuroscience. 1988 Feb;24(2):593–603. doi: 10.1016/0306-4522(88)90353-3. [DOI] [PubMed] [Google Scholar]
  34. Wanke E., Ferroni A., Gattanini P., Meldolesi J. alpha Latrotoxin of the black widow spider venom opens a small, non-closing cation channel. Biochem Biophys Res Commun. 1986 Jan 14;134(1):320–325. doi: 10.1016/0006-291x(86)90565-6. [DOI] [PubMed] [Google Scholar]
  35. Zhu P. C., Thureson-Klein A., Klein R. L. Exocytosis from large dense cored vesicles outside the active synaptic zones of terminals within the trigeminal subnucleus caudalis: a possible mechanism for neuropeptide release. Neuroscience. 1986 Sep;19(1):43–54. doi: 10.1016/0306-4522(86)90004-7. [DOI] [PubMed] [Google Scholar]
  36. de Camilli P., Navone F. Regulated secretory pathways of neurons and their relation to the regulated secretory pathway of endocrine cells. Ann N Y Acad Sci. 1987;493:461–479. doi: 10.1111/j.1749-6632.1987.tb27231.x. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES