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

First Author  Misawa H Year  1994
Journal  J Neurochem Volume  62
Issue  2 Pages  465-70
PubMed ID  8294908 Mgi Jnum  J:16495
Mgi Id  MGI:64570 Doi  10.1046/j.1471-4159.1994.62020465.x
Citation  Misawa H, et al. (1994) Calcium-independent release of acetylcholine from stable cell lines expressing mouse choline acetyltransferase cDNA. J Neurochem 62(2):465-70
abstractText  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 1 alpha 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 Ca(2+)-independent fashion. Neither high K+ nor calcium ionophore stimulated release of ACh from the cells.
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