Calcium‐Independent Release of Acetylcholine from Stable Cell Lines Expressing Mouse Choline Acetyltransferase cDNA

Hidemi Misawa; Ryosuke Takahashi; Takeo Deguchi

doi:10.1046/j.1471-4159.1994.62020465.x

Calcium‐Independent Release of Acetylcholine from Stable Cell Lines Expressing Mouse Choline Acetyltransferase cDNA

Hidemi Misawa, Ryosuke Takahashi, Takeo Deguchi

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

Abstract

Abstract: Stably transfected cells expressing mouse choline acetyltransferase (ChAT) cDNA were established, and the synthesis and release of acetylcholine (ACh) were examined. A cDNA clone coding for mouse ChAT was inserted into an expression vector (pEF321) containing a promoter for human elongation factor 1α to construct pEFmChAT. Neuronal (NG108‐15, NS20Y, N1E115, and Neuro2A) and nonneuronal cell lines (L cells and NIH3T3) were transfected with pEFmChAT, and the cell lines that stably expressed high ChAT activity were selected. These cells expressed the 66‐kDa ChAT protein and accumulated ACh mostly in the cytosol. The concentration of intracellular ACh in the cells increased upon raising the choline level in the medium. The cells continuously released ACh in a Ca²⁺‐independent fashion. Neither high K⁺ nor calcium ionophore stimulated release of ACh from the cells.

Original language	English
Pages (from-to)	465-470
Number of pages	6
Journal	Journal of Neurochemistry
Volume	62
Issue number	2
DOIs	https://doi.org/10.1046/j.1471-4159.1994.62020465.x
Publication status	Published - 1994 Feb
Externally published	Yes

Keywords

Acetylcholine release
Choline acetyltransferase
DNA transfection
Stable transformant

ASJC Scopus subject areas

Biochemistry
Cellular and Molecular Neuroscience

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10.1046/j.1471-4159.1994.62020465.x

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title = "Calcium‐Independent Release of Acetylcholine from Stable Cell Lines Expressing Mouse Choline Acetyltransferase cDNA",

abstract = "Abstract: Stably transfected cells expressing mouse choline acetyltransferase (ChAT) cDNA were established, and the synthesis and release of acetylcholine (ACh) were examined. A cDNA clone coding for mouse ChAT was inserted into an expression vector (pEF321) containing a promoter for human elongation factor 1α to construct pEFmChAT. Neuronal (NG108‐15, NS20Y, N1E115, and Neuro2A) and nonneuronal cell lines (L cells and NIH3T3) were transfected with pEFmChAT, and the cell lines that stably expressed high ChAT activity were selected. These cells expressed the 66‐kDa ChAT protein and accumulated ACh mostly in the cytosol. The concentration of intracellular ACh in the cells increased upon raising the choline level in the medium. The cells continuously released ACh in a Ca2+‐independent fashion. Neither high K+ nor calcium ionophore stimulated release of ACh from the cells.",

keywords = "Acetylcholine release, Choline acetyltransferase, DNA transfection, Stable transformant",

author = "Hidemi Misawa and Ryosuke Takahashi and Takeo Deguchi",

year = "1994",

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doi = "10.1046/j.1471-4159.1994.62020465.x",

language = "English",

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journal = "Journal of Neurochemistry",

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T1 - Calcium‐Independent Release of Acetylcholine from Stable Cell Lines Expressing Mouse Choline Acetyltransferase cDNA

AU - Misawa, Hidemi

AU - Takahashi, Ryosuke

AU - Deguchi, Takeo

PY - 1994/2

Y1 - 1994/2

N2 - Abstract: Stably transfected cells expressing mouse choline acetyltransferase (ChAT) cDNA were established, and the synthesis and release of acetylcholine (ACh) were examined. A cDNA clone coding for mouse ChAT was inserted into an expression vector (pEF321) containing a promoter for human elongation factor 1α to construct pEFmChAT. Neuronal (NG108‐15, NS20Y, N1E115, and Neuro2A) and nonneuronal cell lines (L cells and NIH3T3) were transfected with pEFmChAT, and the cell lines that stably expressed high ChAT activity were selected. These cells expressed the 66‐kDa ChAT protein and accumulated ACh mostly in the cytosol. The concentration of intracellular ACh in the cells increased upon raising the choline level in the medium. The cells continuously released ACh in a Ca2+‐independent fashion. Neither high K+ nor calcium ionophore stimulated release of ACh from the cells.

AB - Abstract: Stably transfected cells expressing mouse choline acetyltransferase (ChAT) cDNA were established, and the synthesis and release of acetylcholine (ACh) were examined. A cDNA clone coding for mouse ChAT was inserted into an expression vector (pEF321) containing a promoter for human elongation factor 1α to construct pEFmChAT. Neuronal (NG108‐15, NS20Y, N1E115, and Neuro2A) and nonneuronal cell lines (L cells and NIH3T3) were transfected with pEFmChAT, and the cell lines that stably expressed high ChAT activity were selected. These cells expressed the 66‐kDa ChAT protein and accumulated ACh mostly in the cytosol. The concentration of intracellular ACh in the cells increased upon raising the choline level in the medium. The cells continuously released ACh in a Ca2+‐independent fashion. Neither high K+ nor calcium ionophore stimulated release of ACh from the cells.

KW - Acetylcholine release

KW - Choline acetyltransferase

KW - DNA transfection

KW - Stable transformant

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DO - 10.1046/j.1471-4159.1994.62020465.x

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AN - SCOPUS:0028144354

SN - 0022-3042

VL - 62

SP - 465

EP - 470

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

IS - 2

ER -

Calcium‐Independent Release of Acetylcholine from Stable Cell Lines Expressing Mouse Choline Acetyltransferase cDNA

Abstract

Keywords

ASJC Scopus subject areas

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