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

Hidemi Misawa, Ryosuke Takahashi, Takeo Deguchi

研究成果: Article査読

18 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)465-470
ページ数6
ジャーナルJournal of Neurochemistry
62
2
DOI
出版ステータスPublished - 1994 2月
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 細胞および分子神経科学

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