Stably transfected cell line that allows for real time tracking of vesicular monoamine transporter 2 (VMAT2) function through the use of fluorescent markers in live cells.
- Can test target compound interaction with human dopamine transporter (hDAT) or VMAT2 in cultured cells instead of in harvested research animal tissue saving time and money.
- Uses fluorescent dyes in assays instead of radioactive markers, which eliminates the need for radiation certification.
- Can test potential VMAT2 agonists or antagonists in a whole cell model as opposed to isolated vesicle assays.
Malfunction of the vesicular monoamine transporter 2 (VMAT2) in dopamine neurons plays a role in Parkinson’s disease, Huntington’s disease, drug abuse, and other neurological diseases. Some environmental toxins, such as certain pesticides, may disrupt VMAT2 function in dopamine neurons. This involvement makes the VMAT2 an attractive therapeutic target for a wide range of health issues. However, current techniques for testing VMAT2 activity involves isolated vesicles from mice and radioactive uptake assays that do not address cell permeability of any test compounds. Therefore, these tests require added research animal and radioactive material costs, training, and certification. By using the DAT/VMAT2 cell line, VMAT2 activity can be assessed using fluorescent markers, while simultaneously confirming cell permeability, thereby decreasing experimental time and costs.
Human embryonic kidney (HEK293) cells were stably transfected with hDAT and the human vesicular monoamine transporter 2 conjugated with an mCherry fluorescent tag. Transfecting these proteins into the HEK293 cells allows for dopamine uptake into the cytosol of the cells and packaging of cytosolic dopamine into vesicles. The fluorescent tag on the VMAT2 allows for validation of vesicular packaging and confirmation of co-localization through fluorescent confocal microscopy.
Cell line has been stably transfected with hDAT and VMAT2 and validation data are available.
Publication: Bernstein et al., J Neurosci Methods, 2012. 209:357-366.