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Free keywords:
ChAT; GFP; Cholinergic marker; In vivo visualization; Mouse (C57/B16); Cell culture; Rat (Wistar)
Abstract:
Choline acetyltransferase (ChAT) is widely used as a marker enzyme to identify cholinergic neurons in the central and peripheral nervous system and to study developmental changes. In order to visualize expression of ChAT directly we have generated a ChAT-green fluorescent protein (GFP) fusion construct. Upon transfection of COS-1 cells and cultured rat hippocampal neurons, transgenic enzymatically active ChAT-GFP is expressed and shows intrinsic fluorescence. In COS-1 cells the ChAT-GFP construct revealed a subcellular distribution indistinguishable from wild-type ChAT. In primary neurons the fluorescence was present in the soma and neuritic processes. Hence, this construct will be useful for analyzing the expression and subcellular distribution of ChAT-GFP in cell and tissue culture.