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Modification of choline acetyltransferase by integration of green fluorescent protein does not affect enzyme activity and subcellular distribution

MPG-Autoren
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Rathenberg,  Jan
Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society;

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Koenen,  Michael
Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society;
Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society;

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Witzemann,  Veit
Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society;
Department of Cell Physiology, Max Planck Institute for Medical Research, Max Planck Society;

Externe Ressourcen

http://link.springer.com/content/pdf/10.1007%2Fs00441-002-0543-x.pdf
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http://dx.doi.org/10.1007/s00441-002-0543-x
(beliebiger Volltext)

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Zitation

Rathenberg, J., Gärtner, A., Koenen, M., & Witzemann, V. (2002). Modification of choline acetyltransferase by integration of green fluorescent protein does not affect enzyme activity and subcellular distribution. Cell and Tissue Research, 308(1), 1-6. doi:10.1007/s00441-002-0543-x.


Zitierlink: https://hdl.handle.net/11858/00-001M-0000-0029-783D-0
Zusammenfassung
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.