Calcium-independent release of acetylcholine from stable cell lines expressing mouse choline acetyltransferase cDNA

Hidemi Misawa, Ryosuke Takahashi, Takeo Deguchi

Research output: Contribution to journalArticle

17 Citations (Scopus)

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 languageEnglish
Pages (from-to)465-470
Number of pages6
JournalJournal of Neurochemistry
Volume62
Issue number2
Publication statusPublished - 1994 Feb
Externally publishedYes

Fingerprint

Choline O-Acetyltransferase
Acetylcholine
Complementary DNA
Cells
Calcium
Cell Line
L Cells (Cell Line)
Peptide Elongation Factor 1
Calcium Ionophores
Choline
Cytosol
Clone Cells
Proteins

Keywords

  • Acetylcholine release
  • Choline acetyltransferase
  • DNA transfection
  • Stable transformant

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Calcium-independent release of acetylcholine from stable cell lines expressing mouse choline acetyltransferase cDNA. / Misawa, Hidemi; Takahashi, Ryosuke; Deguchi, Takeo.

In: Journal of Neurochemistry, Vol. 62, No. 2, 02.1994, p. 465-470.

Research output: Contribution to journalArticle

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