Cell lines expressing an acetylcholine release mechanism; correction of a release-deficient cell by mediatophore transfection.
Several neuronal and non-neuronal cell lines express a Ca(2+)-dependent mechanism of transmitter release that can be demonstrated after loading the cells with acetylcholine during culture. In contrast, a particular cell line, the neuroblastoma N18TG-2, was found to be deficient for release. We transfected N18TG-2 cells with a plasmid encoding Torpedo mediatophore, a protein able to translocate acetylcholine in response to calcium. The N18TG-2 cells expressed the Torpedo protein which reached their plasma membrane. At the same time, these cells acquired a Ca(2+)-dependent quantal release mechanism similar to the one naturally expressed by other cell lines. Hence, the presence of mediatophore in the plasma membrane seems essential for quantal release.
Falk-Vairant
J
J
Départment de Pharmacologie, C.M.U., Genève, Switzerland.
Meunier
F M
FM
Lesbats
B
B
Corrèges
P
P
Eder-Colli
L
L
Salem
N
N
Synguelakis
M
M
Dunant
Y
Y
Israël
M
M
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000378:metabolism; D002460:Cell Line; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004594:Electrophysiology; D006801:Humans; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism; D018408:Patch-Clamp Techniques; D014101:Torpedo; D014162:Transfection; D014407:Tumor Cells, Cultured; D014981:Xenopus
D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; C050299:mediatophore; D000109:Acetylcholine