Monitoring of Black Widow Spider Venom (BWSV) induced exo- and endocytosis in living frog motor nerve terminals with FM1-43

Monitoring of Black Widow Spider Venom (BWSV) induced exo- and endocytosis in living frog motor... The neurotoxin Black Widow Spider Venom (BWSV) triggers massive release of neurotransmitter at synapses. Here we demonstrate that the action of BWSV on the frog neuromuscular junction can be visualized in vivo by the use of the fluorescent styryl dye FM1-43. This vital dye stains recycled synaptic vesicles upon nerve stimulation. Motor nerve terminals were stained with FM1-43 via electrical stimulation, washed and then exposed to BWSV or α-Latrotoxin. All terminals destained completely, independent of external calcium. Exposure of frog nerve terminals to BWSV in the presence of FM1-43 and calcium led to staining of terminals. The staining pattern appeared to be exactly the same as in control preparations, stimulated electrically via the nerve. When the same experiment was performed in the absence of calcium, only a minute quantity of dye was taken up into the nerve terminals and the synapses looked swollen and puffed. Addition of external calcium to these preparations elicited an immediate shrinking of the nerve terminals, indicating endocytosis. These observations support electron-microscopic data that suggest an important role for extracellular calcium in endocytosis of BWSV poisoned nerve terminals. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Neuropharmacology Elsevier

Monitoring of Black Widow Spider Venom (BWSV) induced exo- and endocytosis in living frog motor nerve terminals with FM1-43

Neuropharmacology, Volume 34 (11) – Nov 1, 1995

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Publisher
Elsevier
Copyright
Copyright © 1995 Elsevier Ltd
ISSN
0028-3908
eISSN
1873-7064
D.O.I.
10.1016/0028-3908(95)00126-Q
Publisher site
See Article on Publisher Site

Abstract

The neurotoxin Black Widow Spider Venom (BWSV) triggers massive release of neurotransmitter at synapses. Here we demonstrate that the action of BWSV on the frog neuromuscular junction can be visualized in vivo by the use of the fluorescent styryl dye FM1-43. This vital dye stains recycled synaptic vesicles upon nerve stimulation. Motor nerve terminals were stained with FM1-43 via electrical stimulation, washed and then exposed to BWSV or α-Latrotoxin. All terminals destained completely, independent of external calcium. Exposure of frog nerve terminals to BWSV in the presence of FM1-43 and calcium led to staining of terminals. The staining pattern appeared to be exactly the same as in control preparations, stimulated electrically via the nerve. When the same experiment was performed in the absence of calcium, only a minute quantity of dye was taken up into the nerve terminals and the synapses looked swollen and puffed. Addition of external calcium to these preparations elicited an immediate shrinking of the nerve terminals, indicating endocytosis. These observations support electron-microscopic data that suggest an important role for extracellular calcium in endocytosis of BWSV poisoned nerve terminals.

Journal

NeuropharmacologyElsevier

Published: Nov 1, 1995

References

  • Okadaic acid disrupts clusters of synaptic vesicles in frog motor nerve terminals
    Betz, W.J.; Henkel, A.W.
  • Acetylcholine mobilization in a sympathetic ganglion in the presence and absence of 2-(4-phenylpiperidino)cyclohexanol (AH5183)
    Cabeza, R.; Collier, B.
  • Reversibility and mode of action of Black Widow spider venom on the vertebrate neuromuscular junction
    Gorio, A.; Mauro, A.
  • Redistribution of synaptophysin and synapsin I during alpha-latrotoxin-induced release of neurotransmitter at the neuromuscular junction
    Torri-Tarelli, F.; Villa, A.; Valtorta, F.; De Camilli, .; Greengard, P.; Ceccarelli, B.
  • Specific localization of the alpha-Latrotoxin receptor in the nerve terminal plasma membrane
    Valtorta, F.; Madeddu, L.; Meldolesi, J.; Ceccarelli, B.
  • Synaptophysin (p38) at the frog neuromuscular junction: its incorporation into the axolemma and recycling after intense quantal secretion
    Valtorta, F.; Jahn, R.; Fesce, R.; Greengard, P.; Ceccarelli, B.

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