Discrete acetylcholine release from neuroblastoma or hybrid cells overexpressing choline acetyltransferase into the neuromuscular synaptic cleft

Zhen Guo Zhong, Yasuhiro Kimura, Mami Noda, Hidemi Misawa, Haruhiro Higashida

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Neuroblastoma (clones NS-20Y, N1E-115, and Neuro2A) and neuroblastoma × 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.

Original languageEnglish
Pages (from-to)81-88
Number of pages8
JournalNeuroscience Research
Volume22
Issue number1
DOIs
Publication statusPublished - Jan 1 1995
Externally publishedYes

Fingerprint

Choline O-Acetyltransferase
Hybrid Cells
Neuroblastoma
Acetylcholine
Skeletal Muscle Fibers
Miniature Postsynaptic Potentials
Synapses
Complementary DNA
Transformed Cell Line
Excitatory Postsynaptic Potentials
Coculture Techniques
Glioma
Membrane Potentials
Action Potentials
Transfection
Clone Cells
DNA

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Discrete acetylcholine release from neuroblastoma or hybrid cells overexpressing choline acetyltransferase into the neuromuscular synaptic cleft. / Zhong, Zhen Guo; Kimura, Yasuhiro; Noda, Mami; Misawa, Hidemi; Higashida, Haruhiro.

In: Neuroscience Research, Vol. 22, No. 1, 01.01.1995, p. 81-88.

Research output: Contribution to journalArticle

@article{7f980044e438431481f8e2850cd360bb,
title = "Discrete acetylcholine release from neuroblastoma or hybrid cells overexpressing choline acetyltransferase into the neuromuscular synaptic cleft",
abstract = "Neuroblastoma (clones NS-20Y, N1E-115, and Neuro2A) and neuroblastoma × 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.",
author = "Zhong, {Zhen Guo} and Yasuhiro Kimura and Mami Noda and Hidemi Misawa and Haruhiro Higashida",
year = "1995",
month = "1",
day = "1",
doi = "10.1016/0168-0102(95)00881-S",
language = "English",
volume = "22",
pages = "81--88",
journal = "Neuroscience Research",
issn = "0168-0102",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

TY - JOUR

T1 - Discrete acetylcholine release from neuroblastoma or hybrid cells overexpressing choline acetyltransferase into the neuromuscular synaptic cleft

AU - Zhong, Zhen Guo

AU - Kimura, Yasuhiro

AU - Noda, Mami

AU - Misawa, Hidemi

AU - Higashida, Haruhiro

PY - 1995/1/1

Y1 - 1995/1/1

N2 - Neuroblastoma (clones NS-20Y, N1E-115, and Neuro2A) and neuroblastoma × 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.

AB - Neuroblastoma (clones NS-20Y, N1E-115, and Neuro2A) and neuroblastoma × 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.

UR - http://www.scopus.com/inward/record.url?scp=0028952278&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028952278&partnerID=8YFLogxK

U2 - 10.1016/0168-0102(95)00881-S

DO - 10.1016/0168-0102(95)00881-S

M3 - Article

VL - 22

SP - 81

EP - 88

JO - Neuroscience Research

JF - Neuroscience Research

SN - 0168-0102

IS - 1

ER -