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Evoked acetylcholine release expressed in neuroblastoma cells by transfection of mediatophore cDNA.

Abstract
Transmitter release was elicited in two ways from cultured cells filled with acetylcholine: (a) in a biochemical assay by successive addition of a calcium ionophore and calcium and (b) electrophysiologically, by electrical stimulation of individual cells and real-time recording with an embryonic Xenopus myocyte. Glioma C6-Bu-1 cells were found to be competent for Ca(2+)-dependent and quantal release. In contrast, no release could be elicited from mouse neuroblastoma N18TG-2 cells. However, acetylcholine release could be restored when N18TG-2 cells were transfected with a plasmid coding for mediatophore. Mediatophore is a protein of nerve terminal membranes purified from the Torpedo electric organ on the basis of its acetylcholine-releasing capacity. The transfected N18TG-2 cells expressed Torpedo mediatophore in their plasma membrane. In response to an electrical stimulus, they generated in the myocyte evoked currents that were curare sensitive and calcium dependent and displayed, discrete amplitude levels, like in naturally occurring synapses.
AuthorsJ Falk-Vairant, P Corrèges, L Eder-Colli, N Salem, F M Meunier, B Lesbats, F Loctin, M Synguelakis, M Israël, Y Dunant
JournalJournal of neurochemistry (J Neurochem) Vol. 66 Issue 3 Pg. 1322-5 (Mar 1996) ISSN: 0022-3042 [Print] England
PMID8769901 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • DNA, Complementary
  • Nerve Tissue Proteins
  • mediatophore
  • Acetylcholine
  • Calcium
Topics
  • Acetylcholine (metabolism)
  • Animals
  • Calcium (physiology)
  • DNA, Complementary
  • Electric Stimulation
  • Electrophysiology
  • Glioma (metabolism, pathology)
  • Mice
  • Nerve Tissue Proteins (genetics)
  • Neuroblastoma (metabolism, pathology)
  • Rats
  • Torpedo (metabolism)
  • Transfection
  • Tumor Cells, Cultured
  • Xenopus (embryology)

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