Discrete acetylcholine release from neuroblastoma or hybrid cells overexpressing choline acetyltransferase into the neuromuscular synaptic cleft.
Neuroblastoma (clones NS-20Y, N1E-115, and Neuro2A) and neuroblastoma x glioma hybrid (NG108-15) cells were transfected with mouse choline acetyltransferase (ChAT) complementary DNA (cDNA) or vector DNA alone and stably transformed cell lines were established to examine their ability to secrete acetylcholine (ACh). Membrane potentials were recorded from either presynaptic neuroblastoma and hybrid cells or postsynaptic myotubes in co-culture. After transformation with ChAT, synapses were formed and miniature end-plate potentials (MEPPs) were recorded in myotubes co-cultured with Neuro2A and N1E-115 cells, while parental and mock-transfected control cells totally lacked this ability. The rate of synapse formation and/or MEPP frequency was higher in transformed NG108-15 hybrid and NS-20Y cells than that in the control cells. Action potentials of NS-20Y, Neuro2A or NG108-15 cells overexpressing ChAT were able to evoke end-plate potentials in myotubes, though the average quantum content of these cells was 0.04-0.14, which is as low as the control value. The results show that increased concentrations of ACh by ChAT cDNA transfection reveal a masked property in vesicular ACh release from Neuro2A and N1E-115 cells with no endogenous ChAT activity, or modify their secretory capacity upwardly from NG108-15 and NS-20Y cells with endogenous activity.
Zhong
Z G
ZG
Department of Biophysics, Kanazawa University School of Medicine, Japan.
Kimura
Y
Y
Noda
M
M
Misawa
H
H
Higashida
H
H
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D001932:Brain Neoplasms / Q000201:enzymology / Q000378:metabolism*; D002795:Choline O-Acetyltransferase / Q000096:biosynthesis*; D004558:Electric Stimulation; D006822:Hybrid Cells / Q000201:enzymology / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D009132:Muscles / Q000201:enzymology / Q000294:innervation / Q000378:metabolism; D009447:Neuroblastoma / Q000201:enzymology / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000201:enzymology / Q000378:metabolism* / Q000502:physiology; D051381:Rats; D013569:Synapses / Q000201:enzymology / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000502:physiology
D002795:Choline O-Acetyltransferase; D000109:Acetylcholine