TY - JOUR
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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U2 - 10.1046/j.1471-4159.1994.62020465.x
DO - 10.1046/j.1471-4159.1994.62020465.x
M3 - Article
C2 - 8294908
AN - SCOPUS:0028144354
VL - 62
SP - 465
EP - 470
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
SN - 0022-3042
IS - 2
ER -