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Modification of choline acetyltransferase by integration of green fluorescent protein does not affect enzyme activity
Jan Rathenberg1, Annette Gärtner, Michael Koenen
1Abteilung Zellphysiologie, Max-Planck-Institut für Medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.
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.